LOCUS pEX-N-His 4546 bp DNA circular SYN 12-NOV-2024
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4546)
AUTHORS Ming
TITLE Direct Submission
JOURNAL Exported Nov 12, 2024 from SnapGene 6.0.2 to Vector NTI(R) format
https://www.biofeng.com
COMMENT VNTDATE|1073779200|
COMMENT VNTDBDATE|1073779200|
COMMENT VNTNAME|pEX-N-His|
COMMENT VNTAUTHORNAME|Ming|
FEATURES Location/Qualifiers
primer_bind 166..182
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 218..236
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 237..261
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
RBS 265..287
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 298..315
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
CDS 316..336
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
terminator 462..508
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 754..831
/gene="lacI"
/label=lacI promoter
CDS 832..1914
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter 1963..1993
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2001..2017
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 2025..2041
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2057..2073)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(2365..2953)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3124..3984)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
promoter complement(3985..4089)
/gene="bla"
/label=AmpR promoter
rep_origin 4116..4544
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
181 gtgccgaatt ctgctggact ttccaaaatg tcgagcttaa tacgactcac tataggggaa
241 ttgtgagcgg ataacaattc cccttttgtt taactttaag aaggagaggc caccatgcat
301 caccatcacc atcacgaaaa cctgtatttt cagggcgcga tcgccggcgc gccagatctc
361 aagcttaact agctagcgga ccgacgcgtt aaacgcggcc gctcgaggtt taaacggccg
421 gccacatgac tgactgagcc acgataaccg ctgagcaata actagcataa ccccttgggg
481 cctctaaacg ggtcttgagg ggttttttcc caccagcctt gtcctaatcc gcggcggttc
541 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
601 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
661 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
721 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
781 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
841 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
901 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
961 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
1021 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
1081 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
1141 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgagtcag tgggctgatc
1201 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
1261 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
1321 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
1381 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
1441 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
1501 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
1561 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
1621 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
1681 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
1741 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
1801 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
1861 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
1921 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
1981 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
2041 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
2101 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
2161 aggcccgcac cgatggatcc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
2221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
2281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
2341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
2401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
2461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
2521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
2581 cagttccgtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
2641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
2701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
2761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
2821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
2881 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
2941 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
3001 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
3061 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
3121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
3181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
3241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
3301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
3361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
3421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
3481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
3541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
3601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
3661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
3721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
3781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
3841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
3901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
3961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
4021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
4081 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
4141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
4201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
4261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
4321 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
4381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
4441 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
4501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aatttt
//