LOCUS pEN-L1-NF-L2 3312 bp DNA circular 06-SEP-2022 DEFINITION . ACCESSION urn.local...ey-dt3bdlp KEYWORDS . SOURCE ORGANISM . COMMENT This file is created by Vector NTI http://www.invitrogen.com/ ORIGDB|GenBank VNTDATE|459291304| VNTDBDATE|459291304| LSOWNER| VNTNAME|pEN-L1-NF-L2,0| VNTAUTHORNAME|Demo User| VNTAUTHOREML|manka@psb.ugent.be| Vector_NTI_Display_Data_(Do_Not_Edit!) 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#3=(CRSite (CStringList) "AvaI" "cycgrg" 2 0 1 2427 0 0 "") #4=(CRSite (CStringList) "BamHI" "ggatcc" 2 0 0 0 "") #5=(CRSite (CStringList) "ClaI" "atcgat" 3 0 0 0 "") #6=(CRSite (CStringList) "EcoRI" "gaattc" 2 0 0 0 "") #7=(CRSite (CStringList) "EcoRV" "gatatc" 4 0 1 104 0 0 "") #8=(CRSite (CStringList) "HindIII" "aagctt" 2 0 0 0 "") #9=(CRSite (CStringList) "NcoI" "ccatgg" 2 0 0 0 "") #10=(CRSite (CStringList) "NdeI" "catatg" 3 0 0 0 "") #11=(CRSite (CStringList) "NotI" "gcggccgc" 3 0 0 0 "") #12=(CRSite (CStringList) "NruI" "tcgcga" 4 0 1 343 0 0 "") #13=(CRSite (CStringList) "PstI" "ctgcag" 6 0 0 0 "") #14=(CRSite (CStringList) "SacI" "gagctc" 6 0 0 0 "") #15=(CRSite (CStringList) "SacII" "ccgcgg" 5 0 0 0 "") #16=(CRSite (CStringList) "SalI" "gtcgac" 2 0 0 0 "") #17=(CRSite (CStringList) "SmaI" "cccggg" 4 0 0 0 "") #18=(CRSite (CStringList) "SpeI" "actagt" 2 0 0 0 "") #19=(CRSite (CStringList) "SphI" "gcatgc" 6 0 0 0 "") #20=(CRSite (CStringList) "XbaI" "tctaga" 2 0 0 0 "") #21=(CRSite (CStringList) "XhoI" "ctcgag" 2 0 0 0 "") #22=(CRSite (CStringList) "XmaI" "cccggg" 2 0 0 0 "") #23=(CRSite (CStringList) "BglII" "agatct" 2 0 0 0 "")) (CObList #24=(CFSignal (CObList) "attL1" 86 0 0 2436 2531 0 (CStringList) (CStringList) 1 1 1 1 "2436..2531") #25=(CFSignal (CObList) "attL2" 86 0 1 3306 89 0 (CStringList) (CStringList) 1 1 1 1 "complement(3306..89)") #26=(CFSignal (CObList) "Kan(R)" 22 0 0 260 1069 0 (CStringList) (CStringList "/invitrogen=\"1210000\"") 1 1 1 1 "260..1069") #27=(CFSignal (CObList) "NLS" 85 0 0 2532 2584 0 (CStringList) (CStringList) 1 1 1 1 "2532..2584") #28=(CFSignal (CObList) "GFP" 4 0 0 2585 3301 0 (CStringList) (CStringList) 1 1 1 1 "2585..3301")) (CObList) (CObList) (CObList) (CObList) (CObList) (CObList) (CTextView 0 #29=(CGroupPar (CParagraph 0 (0 0) 1 2 0 0 180) (CObjectList #30=(CRefLinePar (CLinePar (CParagraph 0 (0 0) 0 2 0 0 233) "pEN-L1-NF-L2,0" 2) 5 "" 0 4) #31=(CFolderPar (CGroupPar (CParagraph 1 (0 0) 1 1 0 0 178) (CObjectList #32=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "DNA 'pEN-L1-NF-L2,0'" 1) #33=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "attL1-attL2." 1) #34=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "Local object" 1) #35=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "Created: 10/03/07 08:55" 1) #36=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "Last Modified: 10/03/07 08:55" 1) #37=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "length: 3312 bp" 1) #38=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "storage type: Basic" 1) #39=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "form: Circular" 1))) "General Description") #40=(CFolderPar (CGroupPar (CParagraph 2 (0 0) 1 1 0 0 178) (CObjectList #41=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "Original Source Database: GenBank" 1) #42=(CLinePar (CParagraph 0 (0 0) 1 2 1 0 180) "Modification Date in the Original DB: 1-OCT-2007" 1))) "Standard Fields") #43=(CFolderPar (CGroupPar (CParagraph 8 (0 0) 1 2 0 0 178) (CObjectList)) "Annotations") #44=(CFolderPar (CGroupPar (CParagraph 13 (0 0) 1 1 0 0 178) (CObjectList #45=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27955368 (8 0) 1 1 1 0 178) (CObjectList)) "AatII: 0 sites") 5 1 "GACGTC" "CTGCAG") #46=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 21875912 (8 0) 1 1 1 0 178) (CObjectList #47=(CGroupPar (CParagraph 0 (10 #1# 0) 1 2 2 0 180) (CObjectList #48=(CLinePar (CParagraph 0 (1 #1# 1) 1 2 2 0 191) " N1: 2434 " 1))))) "ApaI: 1 site") 5 1 "GGGCCC" "CCCGGG") #49=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27373512 (8 0) 1 1 1 0 178) (CObjectList)) "AseI: 3 sites") 2 4 "ATTAAT" "TAATTA") #50=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27373608 (8 0) 1 1 1 0 178) (CObjectList #51=(CGroupPar (CParagraph 0 (10 #3# 0) 1 2 2 0 180) (CObjectList #52=(CLinePar (CParagraph 0 (1 #3# 1) 1 2 2 0 191) " N1: 2427 " 1))))) "AvaI: 1 site") 1 5 "CYCGRG" "GRGCYC") #53=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27415288 (8 0) 1 1 1 0 178) (CObjectList)) "BamHI: 0 sites") 1 5 "GGATCC" "CCTAGG") #54=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 22086880 (8 0) 1 1 1 0 178) (CObjectList)) "BglII: 0 sites") 1 5 "AGATCT" "TCTAGA") #55=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27415384 (8 0) 1 1 1 0 178) (CObjectList)) "ClaI: 0 sites") 2 4 "ATCGAT" "TAGCTA") #56=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 28265944 (8 0) 1 1 1 0 178) (CObjectList)) "EcoRI: 0 sites") 1 5 "GAATTC" "CTTAAG") #57=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 28266128 (8 0) 1 1 1 0 178) (CObjectList #58=(CGroupPar (CParagraph 0 (10 #7# 0) 1 2 2 0 180) (CObjectList #59=(CLinePar (CParagraph 0 (1 #7# 1) 1 2 2 0 191) " N1: 104 " 1))))) "EcoRV: 1 site") 3 3 "GATATC" "CTATAG") #60=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 21768048 (8 0) 1 1 1 0 178) (CObjectList)) "HindIII: 0 sites") 1 5 "AAGCTT" "TTCGAA") #61=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 22060400 (8 0) 1 1 1 0 178) (CObjectList)) "NcoI: 0 sites") 1 5 "CCATGG" "GGTACC") #62=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27567672 (8 0) 1 1 1 0 178) (CObjectList)) "NdeI: 0 sites") 2 4 "CATATG" "GTATAC") #63=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27877792 (8 0) 1 1 1 0 178) (CObjectList)) "NotI: 0 sites") 2 6 "GCGGCCGC" "CGCCGGCG") #64=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 22015976 (8 0) 1 1 1 0 178) (CObjectList #65=(CGroupPar (CParagraph 0 (10 #12# 0) 1 2 2 0 180) (CObjectList #66=(CLinePar (CParagraph 0 (1 #12# 1) 1 2 2 0 191) " N1: 343 " 1))))) "NruI: 1 site") 3 3 "TCGCGA" "AGCGCT") #67=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 22100480 (8 0) 1 1 1 0 178) (CObjectList)) "PstI: 0 sites") 5 1 "CTGCAG" "GACGTC") #68=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27416680 (8 0) 1 1 1 0 178) (CObjectList)) "SacI: 0 sites") 5 1 "GAGCTC" "CTCGAG") #69=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 21869232 (8 0) 1 1 1 0 178) (CObjectList)) "SacII: 0 sites") 4 2 "CCGCGG" "GGCGCC") #70=(CRSFolderPar (CFolderPar (CGroupPar (CParagraph 27868656 (8 0) 1 1 1 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FEATURES Location/Qualifiers attL1 1..100 /Color="#00cccc" /created_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="attL1" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL1" misc_feature 48 /Original_Bases="C" /label="C" /note="Geneious type: Editing History Replacement" NLS 103..150 /modified_by="mamee" /label="c2 NLS" CDS 151..867 /label="GFP" primer_bind 434..453 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="26.0" /Tm="60.1" /annotation_group="6050906027296700a4329177-f8ec-4a0c-a467 -f5782d00a3ce" /created_by="primer3" /Sequence="AGGAGCGCACCATCTTCTTC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NF-L2" /label="P47" attL2 complement(869..968) /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="attL2" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL2" primer complement(1006..1026) /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13R" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R (pCDB-Cas9 - GWM13R)" /label="GWM13R" misc_feature 1079..1080 /Original_Bases="TA" /label="TA" /note="Geneious type: Editing History Replacement" promoter 1087..1137 /created_by="chgai" /X-Transferred_From="pCG034_pKan_Kan-CBE" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG034_pKan_Kan-CBE)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG040_ABE-Nuc-NLS_Kan -reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG043_Kan-ABE-Dest_co ntrol_ccdB)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG046_ABE-LbCas12a-NL S_Kan_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG005_pGGD-pKan-KanR- E)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG006_pGGA-Kan_promot er-B)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG035_pKan_Kan)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG039_ABE-N7-NLS_Kan- reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG041_ABE-SV40-NLS_Ka n-reporter_control)" /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Kan_promoter (pCG054_Kan_CBE_control )" /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (KanR)" /label="kan" misc_feature 1138..1947 /modified_by="mamee" /note="Derived using Geneious Prime 2022.1.1 'Annotate from Database' based on nucleotide similarity" /Transferred_From="KanR" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; KanR" /label="KanR" /note="Geneious type: Bacterial Selection" primer_bind 1163..1182 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="25.9" /Tm="59.8" /annotation_group="6050905850958500d81e9db2-b8e7-413f-85b1 -d92bdfabfa60" /created_by="primer3" /Sequence="CGAGGCCGCGATTAAATTCC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NF-L2" /label="P44" primer_bind 1797..1816 /%GC="60.0" /Hairpin_Tm="40.0" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="60509057310316008db25e3b-3dd8-416d-92b6 -46c21ff1b51f" /created_by="primer3" /Sequence="CGCAGACCGATACCAGGATC" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NF-L2" /label="P42" rep_origin 2094..2682 /created_by="thjac" /modified_by="mamee" /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ColE1/pMB1/pBR322/pUC Ori (high-copy-number ColE1/pMB1/pBR322/pUC origin of replication)" /label="ColE1/pMB1/pBR322/pUC Ori" primer_bind 2582..2601 /%GC="55.0" /Hairpin_Tm="37.3" /Self_Dimer_Tm="None" /Tm="59.7" /annotation_group="60509059089988007e54b95b-e2b8-4691-bd6c -2e4eef13a88b" /created_by="primer3" /Sequence="GGGGAAACGCCTGGTATCTT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NF-L2" /label="P45" primer_bind 3190..3209 /%GC="55.0" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /Tm="60.0" /annotation_group="605090596787140026f69c08-1d23-4dac-9fb3 -79cf6d3c56b9" /created_by="primer3" /Sequence="CTTCCGACTGAGCCTTTCGT" /Mismatches=0 /#_Off-target_Sites=0 /Off-target_Template="pEN-L1-NF-L2" /label="P46" primer 3281..3301 /note="Derived using Geneious Prime 2021.2.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pCDB-Cas9 - GWM13F" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F (pCDB-Cas9 - GWM13F)" /label="GWM13F" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccatgcagcc ttctcttaaa 121 cgcatgaaaa tagagccatc ttctcaacct atggtgagca agggcgagga gctgttcacc 181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg 241 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag 361 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc 421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc 481 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac 541 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac 601 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac 661 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc 721 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa 781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc 841 actctcggca tggacgagct gtacaagtac ccagctttct tgtacaaagt tggcattata 901 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc 961 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt 1021 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat 1081 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg 1141 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta 1201 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg 1261 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat 1321 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc 1381 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga 1441 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg 1501 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc 1561 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg 1621 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat 1681 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac 1741 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca 1801 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta 1861 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt 1921 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag 1981 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt 2041 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 2101 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 2161 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 2221 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 2281 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 2341 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 2401 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 2461 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 2521 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 2581 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 2641 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 2701 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 2761 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 2821 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 2881 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac 2941 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt 3001 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag 3061 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg 3121 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa 3181 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc 3241 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt 3301 aagctcgggc cc //