LOCUS       62056_4665        9566 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 9566)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..351
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     terminator      405..657
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(667..683)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    886..910
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2210..2836
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3268..4338
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      4407..4601
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4945..5085
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5271..5859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5949..6740)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7165..7189
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(7267..7441)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7484..8506)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(8574..9251)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    9442..9463
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9478..9508
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9516..9532
                     /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     9540..9556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattctgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca
       61 gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat
      121 cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat
      181 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
      241 caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct
      301 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggacg
      361 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcgatcgt tcaaacattt
      421 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
      481 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
      541 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
      601 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcaag
      661 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      721 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      781 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca gcttgagctt
      841 ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca ggatatattg
      901 gcgggtaaac ctaagagaaa agagcgttta ttagaataac ggatatttaa aagggcgtga
      961 aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat
     1021 caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg
     1081 cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc
     1141 ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc
     1201 gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat
     1261 gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc
     1321 ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg
     1381 gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc
     1441 ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg
     1501 ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg
     1561 ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg
     1621 agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc
     1681 ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag
     1741 gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag
     1801 gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag
     1861 gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc
     1921 aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg
     1981 ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg
     2041 ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg
     2101 agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg
     2161 ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat
     2221 cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg
     2281 cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc
     2341 ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc
     2401 gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc
     2461 gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc
     2521 ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca
     2581 gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa
     2641 aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct
     2701 tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt
     2761 tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat
     2821 taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg
     2881 ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca
     2941 gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg
     3001 aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg
     3061 cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg
     3121 aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg
     3181 aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac
     3241 tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa
     3301 atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag
     3361 cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga
     3421 atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc
     3481 aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat
     3541 agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc
     3601 gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc
     3661 atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc
     3721 atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt
     3781 gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta
     3841 gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag
     3901 aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta
     3961 aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc
     4021 gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc
     4081 gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa
     4141 gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag
     4201 ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag
     4261 gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc
     4321 gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg
     4381 gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg
     4441 tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca
     4501 cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta
     4561 aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca
     4621 gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct
     4681 tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat
     4741 ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc
     4801 accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4861 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4921 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4981 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5041 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     5101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5701 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct
     5941 aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg
     6001 atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc
     6061 gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca
     6121 ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg
     6181 gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga
     6241 tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc
     6301 agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat
     6361 atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg
     6421 ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga
     6481 ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc
     6541 agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg
     6601 tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga
     6661 gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt
     6721 gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac
     6781 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
     6841 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
     6901 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
     6961 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     7021 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     7081 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     7141 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     7201 tagacaactt aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat
     7261 tcgggggatc tggattttag tactggattt tggttttagg aattagaaat tttattgata
     7321 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
     7381 aaaccctata ggaaccctaa ttcccttatc tgggaactac tcacacatta ttatggagaa
     7441 actcgagctt gtcgatcgac agatccggtc ggcatctact ctatttcttt gccctcggac
     7501 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     7561 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     7621 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     7681 gatagagttg gtcaagacca atgcggagca tatacgcccg gagtcgtggc gatcctgcaa
     7741 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     7801 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     7861 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     7921 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     7981 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     8041 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     8101 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg
     8161 cgaccggttg tagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     8221 gtgcacggcg ggagatgcaa taggtcaggc tctcgctaaa ctccccaatg tcaagcactt
     8281 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     8341 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     8401 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     8461 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatatct cattgccccc
     8521 cgggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg atttcagcgt
     8581 gtcctctcca aatgaaatga acttccttat atagaggaag gtcttgcgaa ggatagtggg
     8641 attgtgcgtc atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg
     8701 tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac
     8761 cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt
     8821 aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga
     8881 atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt
     8941 ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttat
     9001 cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc
     9061 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct
     9121 ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga
     9181 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt
     9241 gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga
     9301 cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct
     9361 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     9421 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     9481 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     9541 aggaaacagc tatgaccatg attacg
//
