LOCUS       62056_22955        6566 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 6566)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             267..1058
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    1248..1272
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        1407..1590
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             1599..2390
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      2394..2646
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        2748..3093
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3104..3820
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     3839..3855
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3889..3905)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4151..4175
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4397..5056)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(5057..5826)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
       61 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
      121 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
      181 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc
      241 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
      301 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
      361 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
      421 acctgtccgg tgccctgaat gaacttcagg acgaggcagc gcggctatcg tggctggcca
      481 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
      541 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
      601 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
      661 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
      721 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
      781 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
      841 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
      901 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
      961 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     1021 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     1081 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     1141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     1201 cggggatctc atgctggagt tcttcgccca cccccatggg caaatcgtgg caggatatat
     1261 tgtggtgtaa acacgcgtca cgtggcgcgc cgatcatgag cggagaatta agggagtcac
     1321 gttatgaccc ccgccgatga cgcgggacaa gccgttttac gtttggaact gacagaaccg
     1381 caacgttgaa ggagccactc agccgcgggt ttctggagtt taatgagcta agcacatacg
     1441 tcagaaacca ttattgcgcg ttcaaaagtc gcctaaggtc actatcagct agcaaatatt
     1501 tcttgtcaaa aatgctccac tgacgttcca taaattcccc tcggtatcca attagagtct
     1561 catattcact ctcaatccaa ataatctgca ccggatctat gattgaacaa gatggattgc
     1621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1741 ttgtcaagac cgacctgtcc ggtgccctga atgaacttca ggacgaggca gcgcggctat
     1801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     1861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     1921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     1981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2101 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagatcgtt
     2401 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta
     2461 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
     2521 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag
     2581 aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac
     2641 tagatcggga ttaataggcc tcgtacgacc ggtaacatgg tggagcacga cactctggtc
     2701 tactccaaaa atgtcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa
     2761 caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc
     2821 gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
     2881 gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
     2941 agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgac
     3001 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
     3061 atataaggaa gttcatttca tttggagagg acaattggag ctcatggtga gcaagggcga
     3121 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3181 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3241 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3301 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3361 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3421 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3481 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3541 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3601 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3661 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3721 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3781 cgccgccggg atcactctcg gcatggacga gctgtacaag caccgcggtg gcggccgctc
     3841 tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga
     3901 cctcgagggg gggcccggta cctcgcgatg actaactagc aaatcaccag tctctctcta
     3961 caaatctatc tctctctatt tttctccaga ataatgtgtg agtagttccc agataaggga
     4021 attagggttc ttatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
     4081 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
     4141 ttaaacgcgt tgacaggata tattggcggg taaactaagt cgctgtatgt gtttgtttgt
     4201 attctgcctc ccagagcctg ataaaaacgg tgaatccgtt agcgaggtgc cgccggcttc
     4261 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     4321 gtatagggcg gcgaggcggc tacagccgat agtctggaac agcgcactta cgggttgctg
     4381 cgcaacccaa gtgctaccgg cgcggcagcg tgacccgtgt cggcggctcc aacggctcgc
     4441 catcgtccag aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc
     4501 attcctccgt ttcggtcaag gctggcaggt ctggttccat gcccggaatg ccgggctggc
     4561 tgggcggctc ctcgccgggg ccggtcggta gttgctgctc gcccggatac agggtcggga
     4621 tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca
     4681 cggcggcact gaacaccgac aggcgcaact ggtcgcgggg ctggccccac gccacgcggt
     4741 cattgaccac gtaggccgac acggtgccgg ggccgttgag cttcacgacg gagatccagc
     4801 gctcggccac caagtccttg actgcgtatt ggaccgtccg caaagaacgt ccgatgagct
     4861 tggaaagtgt cttctggctg accaccacgg cgttctggtg gcccatctgc gccacgaggt
     4921 gatgcagcag cattgccgcc gtgggtttcc tcgcaataag cccggcccac gcctcatgcg
     4981 ctttgcgttc cgtttgcacc cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc
     5041 tggactgcgt ggccatgctt atctccatgc ggtagggtgc cgcacggttg cggcaccatg
     5101 cgcaatcagc tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca
     5161 gtcaattaca gattttcttt aacctacgca atgagctatt gcggggggtg ccgcaatgag
     5221 ctgttgcgta cccccctttt ttaagttgtt gatttttaag tctttcgcat ttcgccctat
     5281 atctagttct ttggtgccca aagaagggca cccctgcggg gttcccccac gccttcggcg
     5341 cggctccccc tccggcaaaa agtggcccct ccggggcttg ttgatcgact gcgcggcctt
     5401 cggccttgcc caaggtggcg ctgccccctt ggaacccccg cactcgccgc cgtgaggctc
     5461 ggggggcagg cgggcgggct tcgccttcga ctgcccccac tcgcataggc ttgggtcgtt
     5521 ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca
     5581 cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag
     5641 aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg
     5701 tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag
     5761 cctgtccatc agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt
     5821 ggccgctcgc ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc
     5881 agggcgaagc ccggagagca agcccgtagg gcgccgcagc cgccgtaggc ggtcacgact
     5941 ttgcgaagca aagtctagtg agtatactca agctagcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaa
//
