pME6032

LOCUS       pME6032                9816 bp DNA     circular SYN 13-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9816)
  AUTHORS   Biofeng
  TITLE     Direct Submission
  JOURNAL   Exported Jul 13, 2023 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1180..1809
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=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)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3377..3571
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4281..4826
                     /direction=RIGHT
                     /label=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.
                     "
     protein_bind    5490..5506
                     /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)."
     promoter        complement(5514..5542)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        5776..5853
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(7821..8471)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             8577..9776
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=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
//