LOCUS       62056_4780        9568 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 9568)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..736
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             743..760
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      792..1044
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1066..1090
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2390..3016
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3448..4518
                     /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      4587..4781
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    5125..5265
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6129..6920)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7345..7369
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(7447..7621)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7664..8686)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(8754..9430)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
ORIGIN
        1 catggtagat ctgactagta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
       61 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      121 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      181 gcccgtgccc tggcccaccc tcgtgaccac cttcacctac ggcgtgcagt gcttcagccg
      241 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      301 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      361 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      421 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      481 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
      541 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      601 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
      661 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat
      721 ggacgagctg tacaaggcta gccaccacca ccaccaccac gtgtgaattg gtgaccagct
      781 cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
      841 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
      901 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
      961 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     1021 cggtgtcatc tatgttacta gatcgggaat taaactatca gtgtttgaca ggatatattg
     1081 gcgggtaaac ctaagagaaa agagcgttta ttagaataac ggatatttaa aagggcgtga
     1141 aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat
     1201 caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg
     1261 cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc
     1321 ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc
     1381 gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat
     1441 gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc
     1501 ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg
     1561 gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc
     1621 ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg
     1681 ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg
     1741 ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg
     1801 agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc
     1861 ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag
     1921 gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag
     1981 gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag
     2041 gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc
     2101 aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg
     2161 ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg
     2221 ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg
     2281 agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg
     2341 ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat
     2401 cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg
     2461 cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc
     2521 ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc
     2581 gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc
     2641 gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc
     2701 ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca
     2761 gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa
     2821 aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct
     2881 tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt
     2941 tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat
     3001 taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg
     3061 ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca
     3121 gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg
     3181 aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg
     3241 cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg
     3301 aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg
     3361 aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac
     3421 tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa
     3481 atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag
     3541 cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga
     3601 atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc
     3661 aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat
     3721 agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc
     3781 gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc
     3841 atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc
     3901 atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt
     3961 gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta
     4021 gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag
     4081 aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta
     4141 aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc
     4201 gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc
     4261 gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa
     4321 gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag
     4381 ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag
     4441 gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc
     4501 gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg
     4561 gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg
     4621 tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca
     4681 cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta
     4741 aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca
     4801 gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct
     4861 tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat
     4921 ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc
     4981 accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5041 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5101 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     5161 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5221 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     5281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct
     6121 aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg
     6181 atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc
     6241 gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca
     6301 ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg
     6361 gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga
     6421 tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc
     6481 agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat
     6541 atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg
     6601 ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga
     6661 ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc
     6721 agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg
     6781 tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga
     6841 gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt
     6901 gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac
     6961 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
     7021 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
     7081 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
     7141 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     7201 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     7261 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     7321 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     7381 tagacaactt aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat
     7441 tcgggggatc tggattttag tactggattt tggttttagg aattagaaat tttattgata
     7501 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
     7561 aaaccctata ggaaccctaa ttcccttatc tgggaactac tcacacatta ttatggagaa
     7621 actcgagctt gtcgatcgac agatccggtc ggcatctact ctatttcttt gccctcggac
     7681 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     7741 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     7801 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     7861 gatagagttg gtcaagacca atgcggagca tatacgcccg gagtcgtggc gatcctgcaa
     7921 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     7981 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     8041 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     8101 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     8161 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     8221 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     8281 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg
     8341 cgaccggttg tagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     8401 gtgcacggcg ggagatgcaa taggtcaggc tctcgctaaa ctccccaatg tcaagcactt
     8461 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     8521 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     8581 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     8641 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttcatatct cattgccccc
     8701 cgggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg atttcagcgt
     8761 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
     8821 gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac
     8881 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     8941 ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg
     9001 taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg
     9061 aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct
     9121 tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgttc
     9181 acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct
     9241 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt
     9301 tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat
     9361 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg
     9421 aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac
     9481 gagagtgtcg tgctccacca tgttgggccc ggcgcgccaa gcttggctgc aggtcgacgg
     9541 atccccggga attctaagag gagtccac
//
