LOCUS pEN-L4-BdEF1a-R1 4649 bp DNA circular 06-SEP-2022
DEFINITION .
ACCESSION urn.local...bx-ej4w6yh
KEYWORDS .
SOURCE
ORGANISM .
COMMENT This file is created by Vector NTI
http://www.invitrogen.com/
ORIGDB|GenBank
VNTDATE|628180943|
VNTDBDATE|628180943|
LSOWNER|
VNTNAME|pEN-L4-BdEF1a-R1|
VNTAUTHORNAME|Mansour Karimi|
VNTOAUTHORNAME|Demo User|
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3.45585 1.9 0.123113 1)
#273=(CLabel
(CWidget 0 (0
0) 1 2 0 0 #189# 39703276 100)
(LOGPEN 0 0 8437824) 1
(LOGFONT 38 14 0 0 400 255 0 0 0 3 2 1 34
"Arial") 1.15807 0.611111 0
"{\\i Pst}I (2088)" "@N (@S)" 5 8.03569
-11.5807 0 -10 2.70647 0.694905 #272#)
#274=(CScratch
(CWidget 2 (1 #13# 2) 1 2 0 0 #161# 0 100)
(LOGPEN 0 8 0) 8 1 3.19667 1.9 0.123113 1)
#275=(CLabel (CWidget 0 (0 0) 1 2 0 0 #189# 0 100)
(LOGPEN 0 0 8437824) 1
(LOGFONT 38 14 0 0 400 255 0 0 0 3 2
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"{\\i Pst}I (2280)" "@N (@S)" 5
3.28352 -17.3711 0 -10 2.70647
0.694905 #274#)) (CObjectList))
#276=(CGroupWidget
(CWidget 2 (10 #14# 0) 1 2
0 0 Nil -1056 100) (CObjectList
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#14# 1) 1 2 0 0 #161# 0 100) (LOGPEN
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#278=(CLabel
(CWidget 0 (0 0) 1 2 0 0 #189# 42344476 100)
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#281=(CGroupWidget
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(LOGPEN 0 8 0) 8 1 4.25907 1.9 0.123113 1)
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#285=(CLabel
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#286=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -2388 100)
(CObjectList) (CObjectList))
#287=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -1604
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(CSeqView 10 5 (CObjectList) (CObList) 1 (CObList))
(CObList) 1952543847 (CStringList) 50357091 4063808 (CObList))).
FEATURES Location/Qualifiers
attL4 1..97
/created_by="mamee"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="attL4"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL4"
misc_feature 98..99
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
promoter 100..2099
/created_by="lapau"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="pBdEF1a.2"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; pBdEF1a.2"
/label="pBdEF1a.2"
primer_bind 547..566
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/Sequence="TCAAAGTCAGATTGCGGCAC"
/Tm="59.1"
/%GC="50.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="0.1"
/modified_by="mamee"
/label="P119"
primer_bind 1313..1332
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/Sequence="GTCTTAAGCCCCAGCCTCTC"
/Tm="59.8"
/%GC="60.0"
/Hairpin_Tm="41.3"
/Self_Dimer_Tm="None"
/modified_by="mamee"
/label="P120"
primer_bind 1933..1952
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/Sequence="TACATCTGTCCGCCCACCTA"
/Tm="60.0"
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="None"
/modified_by="mamee"
/label="P121"
attR1 2104..2260
/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="attR1"
/Transferred_Similarity="99.36%"
/Primary_Match="99.36%; Adjusted-Interval-0:0/0; attR1"
misc_feature 2230
/Original_Bases="T"
/label="T"
/note="Geneious type: Editing History Replacement"
primer complement(2323..2339)
/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"
promoter 2401..2451
/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 2452..3261
/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 2477..2496
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="25.9"
/Tm="59.8"
/annotation_group="62447588864827008bd777b5-f633-46d8-804b
-776acc6c25a8"
/created_by="primer3"
/Sequence="CGAGGCCGCGATTAAATTCC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/label="P44"
primer_bind 3111..3130
/%GC="60.0"
/Hairpin_Tm="40.0"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="62447591166312008eb960fc-2f31-4c39-aeca
-7a3cb6dcc63d"
/created_by="primer3"
/Sequence="CGCAGACCGATACCAGGATC"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/label="P42"
rep_origin 3408..3996
/created_by="thjac"
/modified_by="mamee"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC
origin of replication"
/Transferred_Similarity="99.49%"
/Primary_Match="99.49%; 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"
misc_feature 3555
/Original_Bases="C"
/label="C"
/note="Geneious type: Editing History Replacement"
misc_feature 3800..3801
/Original_Bases="AT"
/label="AT"
/note="Geneious type: Editing History Replacement"
primer_bind 3896..3915
/%GC="55.0"
/Hairpin_Tm="37.3"
/Self_Dimer_Tm="None"
/Tm="59.7"
/annotation_group="6244758943660700e2dff746-fa5a-4341-9f11
-76b54cc1bdfa"
/created_by="primer3"
/Sequence="GGGGAAACGCCTGGTATCTT"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/label="P45"
primer_bind 4504..4523
/%GC="55.0"
/Hairpin_Tm="None"
/Self_Dimer_Tm="None"
/Tm="60.0"
/annotation_group="624475900137820070b4c7a4-e87c-43b2-80b1
-bfcfe2132eae"
/created_by="primer3"
/Sequence="CTTCCGACTGAGCCTTTCGT"
/Mismatches=0
/#_Off-target_Sites=0
/Off-target_Template="pEN-L4-BdEF1a-R1"
/label="P46"
primer 4575..4591
/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"
primer 4595..4610
/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"
misc_feature 4628
/Original_Bases="C"
/label="C"
/note="Geneious type: Editing History Replacement"
ORIGIN
1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
61 tgctttttta taatgccaac tttgtataga aaagttggaa gatttgtgat tgttcagcta
121 catgctcctt gaagcacctt tattctacta taaattagtt cgttggacga gactattaga
181 ccacatgcta gccatgcttg ttcgtagttg tcttcaccgc cattgcaaaa attgtcaata
241 aatatttaga gtgggtggca tcagaaaaac atctctagtg gactctcttc ctatcatagc
301 tactcgggct gtagatagaa cgagggcaca agagttgggt ggcgtaggtt tactcgtgac
361 ctcaactctt ttggctgtgt cttacgtcta agatgggttt ggcatgtgag aaacataggt
421 ctaagcaatt catgttaggg ctgttgcatt gttgttgcat caaccaaatg tccagatagc
481 agttcatgct acatctagtt gaaaaccctc atcattaggc ggaacatgtg ttctttttta
541 gcatagtcaa agtcagattg cggcactcgc tcatccacgg aaagaatttt ccctgtgcag
601 gcatctcgat caaaagacgc aaattaattt ttgaatagcg atataacaat atctaattaa
661 cgtttcttgt tttctgcgaa atgtctttca tcataaaatg agtcatctcg atgagcccaa
721 gtgacatagc ccaacacccc accccaccaa taaaagtgaa gaaaacatgt tgggaaaact
781 ataccaagta aaatacgagt tgttctaaag aaaaagtaaa gtacgagtta gatcgcaccc
841 tgtcctggag tgtggcttga tgatccaact cctagcattg tatccctgtt tttggatgat
901 gtaactatta tttacaatga ataaagaggt gttttactag taaaaaaatc ttgaggggag
961 gagaaaataa tggaggtctt ttttcaaacc gatggactat tatttttagt gaaagagaat
1021 aatattattg gaaaaattat tctatccact tattttatat tggcagaata caaagaatgg
1081 tggggtccac gcggaacttg cggcccccga aacctatcga gggcgcggta cccaagcaag
1141 gaacggagga aacttgcggg gcccgaaacc tagtgataaa aggcatatca tccacacgat
1201 gaagatctga cggaccatat ctcccaccac ggaaagccat cagacgagga tcagacggcc
1261 aggaaggaac cctagcgccc gccggtgcca atataaagcg ccactctctc tcgtcttaag
1321 ccccagcctc tccattcccc tctccctctc gccgccgccg tctccttctc ctactccctt
1381 cgaggtgtgt tgttcatccg tcccgaatcc atccatcccc tcttcagatg tgttgttcat
1441 ggctctaata gctctagatc tgcttgtttg tgttgtttag ctctagatct actcgcgcgc
1501 gcttctctct cgatctcctg tagaacaatt ttggttggtt ttttgtgcat atccatggta
1561 attttgtctg caatatggag gaggctttct aagctcctac gtagcatcga tctttagaat
1621 tccctcggtt tctgtttatt tcttcgcgag ggctctctgt tatctgtagg agtagctgta
1681 agcgcggttc gttacggatt aatcgtcatg cttagttgaa cctatcggtc gaaggatttg
1741 tgtgggttgt cgtgtagaat tgacaccatc tacttactgt actgatatgc cgatctgtag
1801 gatactcttc attacttttg tttactgcta gttgtggtgt agatttagca ttctcaaacc
1861 catgctgtag cgtttctaat attgttacat agatctaccg gtgcctgtta attgtattcg
1921 atcgggcgtt tctacatctg tccgcccacc tagttttata tgtggtaatc aaaattgcgt
1981 tgacttcgtg atgctgtctg tgtactgttt ttaatcgctc ttacttagat gatcaacatg
2041 gtgatggtta cgatttactg ttttctaatc cctgttactt cgatgctgca gtttcagcca
2101 tgacaagttt gtacaaaaaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
2161 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatgcagt
2221 cactatgaac caactactta gatggtatta gtgacctgta gaattcgagc tctagagctg
2281 cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
2341 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
2401 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
2461 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
2521 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
2581 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
2641 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
2701 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
2761 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
2821 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
2881 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
2941 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
3001 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
3061 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
3121 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
3181 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
3241 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
3301 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
3361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
3421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
3481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
3541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
3601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
3661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
3721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
3781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
3841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
3901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
3961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
4021 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
4081 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
4141 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
4201 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
4261 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
4321 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
4381 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
4441 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
4501 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
4561 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
4621 gggcccgagt taacgctacc atggagctc
//