LOCUS       62056_4695        8666 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8666)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..76)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    279..303
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1603..2229
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2666..3730
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3799..3993
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4337..4477
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4663..5251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5341..6132)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6557..6581
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(6659..6833)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6876..7898)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(7988..8347)
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     protein_bind    8542..8563
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8578..8608
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8616..8632
                     /label=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)."
     primer_bind     8640..8656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8666
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggca
       61 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
      121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
      181 ccttcccaac agttgcgcag cctgaatggc gaatgctaga gcagcttgag cttggatcag
      241 attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata ttggcgggta
      301 aacctaagag aaaagagcgt ttattagaat aacggatatt taaaagggcg tgaaaaggtt
      361 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
      421 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
      481 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
      541 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
      601 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
      661 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
      721 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
      781 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
      841 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
      901 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
      961 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
     1021 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
     1081 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
     1141 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
     1201 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
     1261 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
     1321 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
     1381 cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt
     1441 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
     1501 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
     1561 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
     1621 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
     1681 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
     1741 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
     1801 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
     1861 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
     1921 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
     1981 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
     2041 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
     2101 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
     2161 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
     2221 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
     2281 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
     2341 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
     2401 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
     2461 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
     2521 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
     2581 ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc
     2641 cccaagcccg aggaatcggc gtgacggtcg caaaccatcc ggcccggtac aaatcggcgc
     2701 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg
     2761 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa
     2821 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga
     2881 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag
     2941 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat
     3001 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag
     3061 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg
     3121 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt
     3181 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg
     3241 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg
     3301 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga
     3361 aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac
     3421 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg
     3481 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg
     3541 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt
     3601 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc
     3661 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc
     3721 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg
     3781 tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg
     3841 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta
     3901 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat
     3961 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga
     4021 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg
     4081 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg
     4141 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc
     4201 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4261 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4321 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     4381 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4441 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     4501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt ctaggtacta
     5341 aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag
     5401 taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga tcgaccggac
     5461 gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc
     5521 acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac
     5581 aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa
     5641 tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta
     5701 atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat
     5761 ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc
     5821 atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca
     5881 gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag
     5941 catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat
     6001 ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag gagacattcc
     6061 ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct
     6121 cattttagcc atttattatt tccttcctct tttctacagt atttaaagat accccaagaa
     6181 gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac
     6241 cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag
     6301 ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta caatcaacat
     6361 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga
     6421 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt
     6481 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga
     6541 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac
     6601 ttaataacac attgcggacg tttttaatgt actgaattaa cgccgaatta attcggggga
     6661 tctggatttt agtactggat tttggtttta ggaattagaa attttattga tagaagtatt
     6721 ttacaaatac aaatacatac taagggtttc ttatatgctc aacacatgag cgaaacccta
     6781 taggaaccct aattccctta tctgggaact actcacacat tattatggag aaactcgagc
     6841 ttgtcgatcg acagatccgg tcggcatcta ctctatttct ttgccctcgg acgagtgctg
     6901 gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg
     6961 cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg
     7021 catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt
     7081 tggtcaagac caatgcggag catatacgcc cggagtcgtg gcgatcctgc aagctccgga
     7141 tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag
     7201 aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc
     7261 gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg
     7321 tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg
     7381 gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc
     7441 gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg
     7501 aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatagcctc cgcgaccggt
     7561 tgtagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg
     7621 cgggagatgc aataggtcag gctctcgcta aactccccaa tgtcaagcac ttccggaatc
     7681 gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg
     7741 cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat
     7801 tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac
     7861 ttctcgacag acgtcgcggt gagttcaggc tttttcatat ctcattgccc cccgggatct
     7921 gcgaaagctc gagagagata gatttgtaga gagagactgg tgattggtta cttcctaatc
     7981 gaagctttgc ttgggtagaa taggtaagtc agattgaatc tgaaataaag ggaggaaggg
     8041 cgaacttaag aaggtatgac cgggtcgttc acttaccttg cttgacaaac gcaccaagtt
     8101 atcgtgcacc aagcagcaga tgataataat gtcctcgttc ctgtctgcta ataagagtca
     8161 cacttcgagc gccgccgcta ctgctacaag tggggctgat ctgaccagtt gcctaaatga
     8221 accatcttgt caaacgacac aaattttgtg ctcaccgcct ggacgactaa accaaaatag
     8281 gcattcattg ttgacctcca ctagctccag ccaagcccaa aaagtgctcc ttcaatatca
     8341 tcttctgtcg aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga
     8401 cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct
     8461 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     8521 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     8581 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     8641 aggaaacagc tatgaccatg attacg
//
