LOCUS Exported 5315 bp ds-DNA circular SYN 27-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5315
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 141..159
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 160..184
/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 199..221
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 240..257
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 266..315
/label=MCS
CDS 316..333
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 400..447
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 484..939
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(1032..1847)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 1969..2557
/direction=RIGHT
/label=pBR322 ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2743..2885
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2987..3178)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3987..5069)
/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(5070..5147)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
61 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
121 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
181 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
241 atcatcatca tcatcacagc agcggccgcg gatccgaatt cgagctccgt cgacaagctt
301 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
361 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
421 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
481 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
541 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
601 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
661 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
721 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
781 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
841 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
901 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact tttcggggaa
961 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
1021 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
1081 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
1141 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
1201 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
1261 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
1321 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
1381 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
1441 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
1501 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
1561 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
1621 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
1681 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
1741 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
1801 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta
1861 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt
1921 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
1981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
2041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
2101 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
2161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
2221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
2281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
2341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
2401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
2461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
2521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
2581 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
2641 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
2701 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
2761 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc
2821 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
2881 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
2941 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
3001 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
3061 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
3121 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
3181 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
3241 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
3301 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
3361 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
3421 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
3481 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
3541 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
3601 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
3661 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
3721 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
3781 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
3841 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
3901 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
3961 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
4021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
4081 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
4141 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
4201 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
4261 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
4321 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
4381 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
4441 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
4501 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
4561 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
4621 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
4681 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
4741 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
4801 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
4861 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
4921 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
4981 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
5041 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
5101 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
5161 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
5221 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
5281 ggggcctgcc accataccca cgccgaaaca agcgc
//