LOCUS Exported 9816 bp ds-DNA circular SYN 10-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pME6032
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9816)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9816
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1180..1809
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2238..3311
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 RepA"
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 3377..3571
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 4281..4826
/direction=RIGHT
/note="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
misc_feature 5148..5414
/note="T4 terminator"
misc_feature 5416..5473
/note="MCS"
misc_feature 5478..5481
/note="RBS"
protein_bind 5490..5506
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
misc_feature complement(5514..5542)
/note="Tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
promoter 5776..5853
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 5977..6792
/note="LacIQ"
CDS complement(7821..8471)
/codon_start=1
/gene="tetR"
/product="tetracycline resistance regulatory protein"
/note="TetR"
/translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
CDS 8577..9776
/codon_start=1
/gene="tetA"
/product="tetracycline efflux protein"
/note="TcR"
/note="confers resistance to tetracycline"
/translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
"
ORIGIN
1 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
61 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
121 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
181 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
241 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
301 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
361 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
421 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
481 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
541 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
601 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
661 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
721 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
781 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
841 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
901 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
961 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
1021 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
1081 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
1141 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
1201 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
1261 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
1321 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
1381 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
1441 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
1501 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
1561 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
1621 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
1681 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
1741 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
1801 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
1861 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
1921 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
1981 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
2041 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
2101 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
2161 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
2221 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
2281 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
2341 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
2401 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
2461 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
2521 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
2581 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
2641 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
2701 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
2761 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
2821 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
2881 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
2941 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
3001 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
3061 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
3121 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
3181 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
3241 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
3301 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
3361 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
3421 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
3481 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
3541 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
3601 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
3661 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
3721 gcggacaagc cgcgccgtcg ccactcgacc gccggcgctg aggtctgcct cgtgaagaag
3781 gtgttgctga ctcataccag ggcaatggca acaacgttgc gcaaactatt aactggcgaa
3841 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
3901 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
3961 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
4021 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
4081 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
4141 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4201 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4261 cccttaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
4321 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
4381 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
4441 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
4501 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
4561 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
4621 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
4681 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
4741 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
4801 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
4861 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
4921 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
4981 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
5041 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgaggtc
5101 tgcctcgtga agaaggtgtt gctgactcat accaggctag tccgaggagc tttatgcttg
5161 taaaccgttt tgtgaaaaaa tttttaaaat aaaaaagggg acctctaggg tccccaatta
5221 attagtaata taatctatta aaggtcattc aaaaggtcat ccaccggatc aattcccctg
5281 ctcgcgcagg ctgggtgcca agctctcggg taacatcaag gcccgatcct tggagccctt
5341 gccctcccgc acgatgatcg tgccgtgatc gaaatccaga tccttgaccc gcagttgcaa
5401 accctcactg atccgctagt ccgaggcctc gagatctatc gatgcatgcc atggtacccg
5461 ggagctcgaa ttctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacattata
5521 cgagccgatg attaattgtc aacagctcat ttcagaatat ttgccagaac cgttatgatg
5581 tcggcgcaaa aaacattatc cagaacggga gtgcgccttg agcgacacga attatgcagt
5641 gatttacgac ctgcacagcc ataccacagc ttccgatggc tgcctgacgc cagaagcatt
5701 ggtgcaccgt gcagtcgata agcccggatc agcttgcaat tcgcgcgcga aggcgaagcg
5761 gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag
5821 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
5881 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
5941 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
6001 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
6061 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
6121 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
6181 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
6241 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
6301 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
6361 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
6421 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
6481 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
6541 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
6601 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
6661 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
6721 aaacaggatt ttcgcctgct ggggcaaacc acgtggaccg cttgctgcaa ctctctcagg
6781 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
6841 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
6901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
6961 cgcgaattgc aagctggccg acgcgctggg ctacgtcttg ctggcgttcg gatccgatat
7021 cgccgtggcg gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
7081 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
7141 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
7201 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
7261 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
7321 actggcagag cattacgctg acttgacggg acggcggctt tgttgaataa atcgaacttt
7381 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
7441 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
7501 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
7561 ttcacgaggc agacctcagc gccaggcgca cgaggccggc gcgcggaccc gcgccacggc
7621 ccacgagcgg acgccgcagc aggagcgcca gaaggccgcc agagaggccg agcgcggccg
7681 tgaggcttgg acgctagggc agggcatgaa aaagcccgta gcgggctgct acgggcgtct
7741 gacgcggtgg aaagggggag gggatgttgt ctacatggct ctgctgtagt gagtgggttg
7801 cgctccggca gcggtcctga tcaatcgtca ccctttctcg gtccttcaac gttcctgaca
7861 acgagcctcc ttttcgccaa tccatcgaca atcaccgcga gtccctgctc gaacgctgcg
7921 tccggaccgg cttcgtcgaa ggcgtctatc gcggcccgca acagcggcga gagcggagcc
7981 tgttcaacgg tgccgccgcg ctcgccggca tcgctgtcgc cggcctgctc ctcaagcacg
8041 gccccaacag tgaagtagct gattgtcatc agcgcattga cggcgtcccc ggccgaaaaa
8101 cccgcctcgc agaggaagcg aagctgcgcg tcggccgttt ccatctgcgg tgcgcccggt
8161 cgcgtgccgg catggatgcg cgcgccatcg cggtaggcga gcagcgcctg cctgaagctg
8221 cgggcattcc cgatcagaaa tgagcgccag tcgtcgtcgg ctctcggcac cgaatgcgta
8281 tgattctccg ccagcatggc ttcggccagt gcgtcgagca gcgcccgctt gttcctgaag
8341 tgccagtaaa gcgccggctg ctgaaccccc aaccgttccg ccagtttgcg tgtcgtcaga
8401 ccgtctacgc cgacctcgtt caacaggtcc agggcggcac ggatcactgt attcggctgc
8461 aactttgtca tgcttgacac tttatcactg ataaacataa tatgtccacc aacttatcag
8521 tgataaagaa tccgcgcgtt caatcggacc agcggaggct ggtccggagg ccagacgtga
8581 aacccaacat acccctgatc gtaattctga gcactgtcgc gctcgacgct gtcggcatcg
8641 gcctgattat gccggtgctg ccgggcctcc tgcgcgatct ggttcactcg aacgacgtca
8701 ccgcccacta tggcattctg ctggcgctgt atgcgttggt gcaatttgcc tgcgcacctg
8761 tgctgggcgc gctgtcggat cgtttcgggc ggcggccaat cttgctcgtc tcgctggccg
8821 gcgccactgt cgactacgcc atcatggcga cagcgccttt cctttgggtt ctctatatcg
8881 ggcggatcgt ggccggcatc accggggcga ctggggcggt agccggcgct tatattgccg
8941 atatcactga tggcgatgag cgcgcgcggc acttcggctt catgagcgcc tgtttcgggt
9001 tcgggatggt cgcgggacct gtgctcggtg ggctgatggg cggtttctcc ccccacgctc
9061 cgttcttcgc cgcggcagcc ttgaacggcc tcaatttcct gacgggctgt ttccttttgc
9121 cggagtcgca caaaggcgaa cgccggccgt tacgccggga ggctctcaac ccgctcgctt
9181 cgttccggtg ggcccggggc atgaccgtcg tcgccgccct gatggcggtc ttcttcatca
9241 tgcaacttgt cggacaggtg ccggccgcgc tttgggtcat tttcggcgag gatcgctttc
9301 actgggacgc gaccacgatc ggcatttcgc ttgccgcatt tggcattctg cattcactcg
9361 cccaggcaat gatcaccggc cctgtagccg cccggctcgg cgaaaggcgg gcactcatgc
9421 tcggaatgat tgccgacggc acaggctaca tcctgcttgc cttcgcgaca cggggatgga
9481 tggcgttccc gatcatggtc ctgcttgctt cgggtggcat cggaatgccg gcgctgcaag
9541 caatgttgtc caggcaggtg gatgaggaac gtcaggggca gctgcaaggc tcactggcgg
9601 cgctcaccag cctgacctcg atcgtcggac ccctcctctt cacggcgatc tatgcggctt
9661 ctataacaac gtggaacggg tgggcatgga ttgcaggcgc tgccctctac ttgctctgcc
9721 tgccggcgct gcgtcgcggg ctttggagcg gcgcagggca acgagccgat cgctgatcgt
9781 ggaaacgata ggccgctcta gaactagcta gaacta
//