LOCUS Exported 6169 bp ds-DNA circular SYN 17-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..6169
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 59..81
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 101..118
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 128..781
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 818..832
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=EK
/translation="DDDDK"
CDS 893..910
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 977..1024
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1061..1516
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1543..1647
/gene="bla"
/label=AmpR promoter
CDS 1648..2508
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2679..3267
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 3453..3595
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(3697..3888)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4697..5779)
/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 complement(5780..5857)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gtctggttct catcatcatc atcatcatag
121 cagcggtatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
181 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
241 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
301 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
361 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
421 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
481 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
541 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
601 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
661 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
721 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
781 atcggatctg ggccacacag gccatagatc tggtaccgac gacgacgaca aggccatggc
841 tgatatcgga tccgaattcg agctcgtcga caagcttgcg gccgcactcg agcaccacca
901 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
961 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
1021 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
1081 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
1141 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
1201 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
1261 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
1321 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
1381 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
1441 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
1501 ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
1561 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
1621 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
1681 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
1741 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
1801 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
1861 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
1921 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
1981 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
2041 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
2101 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
2161 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
2221 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
2281 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
2341 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
2401 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
2461 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
2521 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
2581 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
2641 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
2701 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
2761 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
2821 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
2881 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
2941 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
3001 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
3061 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
3121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
3181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
3241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
3301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
3361 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
3421 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
3481 ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg
3541 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
3601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
3661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
3721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
3781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
3841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
3901 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
3961 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
4021 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
4081 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
4141 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
4201 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
4261 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
4321 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
4381 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
4441 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
4501 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
4561 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
4621 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
4681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
4801 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
4861 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
4921 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
4981 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
5041 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
5101 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
5161 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
5221 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
5281 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
5341 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
5401 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
5461 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
5521 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
5581 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
5641 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
5701 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
5761 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
5821 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
5881 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
5941 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
6001 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
6061 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
6121 acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaat
//