LOCUS pMG414 6655 bp DNA circular 15-FEB-2023 DEFINITION . ACCESSION urn.local...9i-citr5z2 KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ VNTDATE|630092750| VNTDBDATE|635858458| LSOWNER| VNTNAME|pMG414| VNTAUTHORNAME|Mansour Karimi| VNTOAUTHORNAME|Demo User| Vector_NTI_Display_Data_(Do_Not_Edit!) 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FEATURES Location/Qualifiers attR4 1..124 /created_by="mamee" /Color="#00cccc" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attR4" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attR4" /Alternative_Match="99.19%; Adjusted-Interval-0:0/0; attR4" CDS complement(165..470) /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="ccdB" /Transferred_Similarity="99.35%" /Primary_Match="99.35%; Adjusted-Interval-0:0/0; ccdB" /label="ccdB" primer_bind 425..445 /%GC="47.6" /Hairpin_Tm="45.0" /Self_Dimer_Tm="None" /Tm="57.6" /annotation_group="62457029504545009e7764c7-46fe-4324-80a0 -d54fdf68e792" /created_by="primer3" /Sequence="GACGATAACGGCTCTCTCTTT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P15" UTR complement(471..789) /Mismatches=1 /%_Identity="99.68652037617555" /Motif="AAGCCAGATAACAGTATGCGTATTTGCGCGCTGATTTTTGCGGTATAAGA ATATATACTGATATGTATACCCGAAGTATGTCAAAAGGAGGTATGCTATGAAGCAGCG TATTACAGTGACAGTTGACAGCGACAGCTATCAGTTGCTCAAGGCATATATGATGTCA ATATCTCCGGTCTGGTAAGCACAACCATGCAGAATGAAGCCCGTCGTCTGCGTGCCGA ACGCTGGAAAGCGGAAAATCAGGAAGGGATGGCTGAGGTCGCCCGGTTTATTGAAATG AACGGCTCTTTTGCTGACGAGAACAGGGGCTGGTGAA" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="ccdB" /Transferred_Similarity="98.12%" /Primary_Match="98.12%; Adjusted-Interval-0:0/0; ccdB 5'UTR (ccdB)" /label="ccdB 5'UTR" primer_bind complement(514..533) /%GC="55.0" /Hairpin_Tm="35.1" /Self_Dimer_Tm="None" /Tm="60.1" /annotation_group="62457028853011003d666165-d06e-4d30-bae9 -a0d774c0211d" /created_by="primer3" /Sequence="CTGAGGTCGCCCGGTTTATT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P13" misc_feature complement(812..1470) /Transferred_From="cmR" /Transferred_Similarity="99.85%" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="CmR" /Transferred_Similarity="99.85%" /Primary_Match="99.85%; Adjusted-Interval-0:0/0; CmR" /label="CmR" /note="Geneious type: Bacterial Selection" primer_bind 1221..1240 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6245703007309700d870edfa-3409-47ba-b689 -f64c61fbb756" /created_by="primer3" /Sequence="TCACCAGCTCACCGTCTTTC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P16" promoter complement(1524..1554) /note="Lac promoter of Escherichia coli containing 2 base pair mutations" /regulatory_class="promoter" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /X-Transferred_From="lacUV5" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="CmR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacUV5 (CmR)" /label="lacUV5" attR3 complement(1578..1702) /created_by="mamee" /Color="#00cccc" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attR3" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attR3" terminator 1716..1955 /modified_by="mamee" /label="tCYC1" primer_bind 1869..1888 /%GC="45.0" /Hairpin_Tm="37.1" /Self_Dimer_Tm="25.1" /Tm="57.8" /annotation_group="6245614310067700d8bd9593-5c0c-451a-9247 -f2653cd34e7b" /created_by="primer3" /Sequence="TTTTCTGTACAGACGCGTGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P128" promoter complement(1976..1993) /created_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="T7 promoter" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7 promoter" /label="T7 promoter" primer complement(2004..2019) /invitrogen=1980000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 (-20) forward primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 (-20) forward primer (pDONRP4-P3 - M13 (-20) forward primer)" /label="M13 (-20) forward primer" primer complement(2023..2039) /invitrogen=2000000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 (-40) forward primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 (-40) forward primer (pDONRP4-P3 - M13 (-40) forward primer)" /label="M13 (-40) forward primer" rep_origin 2160..2615 /note="f1 replication origin derived from bacteriophage f1" /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="F1" /Transferred_Similarity="99.56%" /Primary_Match="99.56%; Adjusted-Interval-0:0/0; F1" /label="F1" misc_feature 2159^2160 /Original_Bases="CGCG" /label="CGCG" /note="Geneious type: Editing History Deletion" primer_bind 2343..2361 /%GC="63.2" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.4" /annotation_group="62457032141635004a029b85-9448-45dc-bf1d -801f0c424865" /created_by="primer3" /Sequence="CTTTACGGCACCTCGACCC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P70" misc_feature 2606 /Original_Bases="T" /label="T" /note="Geneious type: Editing History Replacement" misc_feature 2669..2672 /Original_Bases /label /note="Geneious type: Editing History Insertion" CDS complement(2718..3392) /label="TRP1" misc_feature 3633 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 3674..3675 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 3677 /Original_Bases /label /note="Geneious type: Editing History Insertion" primer_bind complement(3817..3836) /%GC="55.0" /Hairpin_Tm="33.5" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6245702732448700257dd51b-1bca-4548-a5a5 -f2ccca5b353c" /created_by="primer3" /Sequence="ACCTCTGACACATGCAGCTC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P5" regulatory complement(3928..4445) /modified_by="mamee" /label="CEN6_ARS4" misc_feature 3928^3929 /Original_Bases="G" /label="G" /note="Geneious type: Editing History Deletion" promoter 4472..4576 /created_by="mamee" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="AmpR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; amp (AmpR)" /label="amp" primer_bind complement(4520..4541) /%GC="50.0" /Hairpin_Tm="None" /Self_Dimer_Tm="18.4" /Tm="60.2" /annotation_group="6245613753549700dbec34cb-35da-4f1c-94ee -0abfe127dd2f" /created_by="primer3" /Sequence="AGGGTTATTGTCTCATGAGCGG" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P21" misc_feature 4577..5437 /X-Transferred_From="AmpR" /X-Transferred_Similarity="100.00%" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="AmpR" /Transferred_Similarity="99.77%" /Primary_Match="99.77%; Adjusted-Interval-0:0/0; AmpR" /label="AmpR" /note="Geneious type: Bacterial Selection" primer_bind complement(5103..5122) /%GC="55.0" /Hairpin_Tm="40.4" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="6245702641730700a0ceb01e-326a-430a-96b7 -c34fadcfe642" /created_by="primer3" /Sequence="TACAGGCATCGTGGTGTCAC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P20" rep_origin 5592..6211 /label="pBR322_origin" misc_feature 5755 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" primer_bind complement(5788..5807) /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="58.9" /annotation_group="62456138692169008a1e0b6b-23ad-4077-b88a -60887165d775" /created_by="primer3" /Sequence="GCGGTGCTACAGAGTTCTTG" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P6" misc_feature 6496 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" misc_feature 6543 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" primer_bind complement(6549..6568) /%GC="50.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="59.9" /annotation_group="6245613625249800295a9ef5-e441-44c9-994e -b8c30b7e2c9c" /created_by="primer3" /Sequence="TCCGCTCACAATTCCACACA" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pMG414" /label="P19" primer 6560..6581 /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pGG-B-HA-TurboID-linker-C - GG_SEQ_fw" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GG_SEQ_fw (pGG-B-HA-TurboID-linker-C - GG_SEQ_fw)" /label="GG_SEQ_fw" primer 6582..6598 /invitrogen=2010000 /note="Derived using Geneious Prime 2020.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pDONRP4-P3 - M13 reverse primer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; M13 reverse primer (pDONRP4-P3 - M13 reverse primer)" /label="M13 reverse primer" ORIGIN 1 caactttgta tagaaaagtt gaacgagaaa cgtaaaatga tataaatatc aatatattaa 61 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac 121 tatggtcgac ctgcagactg gctgtgtata agggagcctg acatttatat tccccagaac 181 atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc 241 ccgataacgg agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc 301 ccgatatgca ccaccgggta aagttcacgg gggactttat ctgacagcag acgtgcactg 361 gccaggggga tcaccatccg tcgcccgggc gtgtcaataa tatcactctg tacatccaca 421 aacagacgat aacggctctc tcttttatag gtgtaaacct taaactgcat ttcaccagcc 481 cctgttctcg tcgacaaaag agccgttcat ttcaataaac cgggcgacct cagccatccc 541 ttcttgattt tccgctttcc agcgttcggc acgcagacga cgggcttcat tctgcatggt 601 tgtgcttacc gaaccggaga tactgacatc atatatgcct tgagcaactg atagctgtcg 661 ctgtcaactg tcactgtaat acgctgcttc atagcatacc tctttttgac atacttcggg 721 tatacatatc agtatatatt cttataccgc aaaaatcagc gcgcaaatac gcatactgtt 781 atctggcttt tagtaagccg gatcctctag attacgcccc gcctgccact catcgcagta 841 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac 901 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa 961 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac 1021 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag 1081 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc 1141 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca 1201 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg 1261 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt 1321 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca 1381 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac 1441 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga 1501 cggatcctaa ctcaaaatcc acacattata cgagccggaa gcataaagtg taaagcctgg 1561 ggtgcctaat gcggccgcca tagtgactgg atatgttgtg ttttacagta ttatgtagtc 1621 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt 1681 tcaactttat tatacatagt tgctcgagtc atgtaattag ttatgtcacg cttacattca 1741 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag 1801 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt 1861 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt 1921 gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata 1981 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 2041 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 2101 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga 2161 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 2221 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 2281 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 2341 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 2401 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 2461 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 2521 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 2581 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg 2641 catctgtgcg gtatttcaca ccgcatagat cggcaagtgc acaaacaata cttaaataaa 2701 tactactcag taataaccta tttcttagca tttttgacga aatttgctat tttgttagag 2761 tcttttacac catttgtctc cacacctccg cttacatcaa caccaataac gccatttaat 2821 ctaagcgcat caccaacatt ttctggcgtc agtccaccag ctaacataaa atgtaagctt 2881 tcggggctct cttgccttcc aacccagtca gaaatcgagt tccaatccaa aagttcacct 2941 gtcccacctg cttctgaatc aaacaaggga ataaacgaat gaggtttctg tgaagctgca 3001 ctgagtagta tgttgcagtc ttttggaaat acgagtcttt taataactgg caaaccgagg 3061 aactcttggt attcttgcca cgactcatct ccatgcagtt ggacgatatc aatgccgtaa 3121 tcattgacca gagccaaaac atcctcctta ggttgattac gaaacacgcc aaccaagtat 3181 ttcggagtgc ctgaactatt tttatatgct tttacaagac ttgaaatttt ccttgcaata 3241 accgggtcaa ttgttctctt tctattgggc acacatataa tacccagcaa gtcagcatcg 3301 gaatctagag cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga 3361 ccagaactac ctgtgaaatt aataacagac atactccaag ctgcctttgt gtgcttaatc 3421 acgtatactc acgtgctcaa tagtcaccaa tgccctccct cttggccctc tccttttctt 3481 ttttcgaccg aattaattct taatcggcaa aaaaagaaaa gctccggatc aagattgtac 3541 gtaaggtgac aagctatttt tcaataaaga atatcttcca ctactgccat ctggcgtcat 3601 aactgcaaag tacacatata ttacgatgct gttctattaa atgcttccta tattatatat 3661 atagtaatgt cgtgatctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt 3721 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc 3781 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 3841 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt 3901 taatgtcatg ataataatgg tttcttagac ggatcgcttg cctgtaactt acacgcgcct 3961 cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt 4021 ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa 4081 aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta 4141 gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc 4201 acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc 4261 tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt 4321 acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta 4381 atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa 4441 ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 4501 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 4561 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 4621 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 4681 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 4741 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 4801 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 4861 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 4921 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 4981 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 5041 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 5101 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 5161 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 5221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 5281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 5341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 5401 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 6301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 6361 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 6421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 6481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 6541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 6601 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctc //