LOCUS       62056_3705        5996 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 5996)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             458..1249
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1657..1678
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1693..1723
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /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     1755..1771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1792..1810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1850..1954
                     /label=AmpR promoter
     CDS             1955..2671
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2672..2737
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2791..2809)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2819..2835)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3545..4204)
                     /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(4205..4974)
                     /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"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
      241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
      301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
      361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
      421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
     1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
     1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1801 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacc gcggaacccc
     1861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1921 ataaatgctt caataatatt gaaaaaggaa gagtatggtg agcaagggcg aggagctgtt
     1981 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2041 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2101 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2161 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2221 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2281 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2341 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2401 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2461 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2521 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2581 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2641 gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc
     2701 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aatctagagc
     2761 ggccgccacc gcggtggagc tccaattcgc cctatagtga gtcgtattac gcgcgctcac
     2821 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2881 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2941 cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt
     3001 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgactgcga
     3061 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     3121 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
     3181 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
     3241 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
     3301 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
     3361 agagcctgat aaaaacggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     3421 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     3481 gaggcggcta cagccgatag tctggaacag cgcacttacg ggttgctgcg caacccaagt
     3541 gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa
     3601 aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt
     3661 cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct
     3721 cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt
     3781 cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga
     3841 acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt
     3901 aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca
     3961 agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct
     4021 tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca
     4081 ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg
     4141 tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg
     4201 ccatgcttat ctccatgcgg tagggtgccg cacggttgcg gcaccatgcg caatcagctg
     4261 caacttttcg gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga
     4321 ttttctttaa cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc
     4381 cccctttttt aagttgttga tttttaagtc tttcgcattt cgccctatat ctagttcttt
     4441 ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc
     4501 cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca
     4561 aggtggcgct gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg
     4621 ggcgggcttc gccttcgact gcccccactc gcataggctt gggtcgttcc aggcgcgtca
     4681 aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc gccttccact tggtgtccaa
     4741 ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa aattaaaaaa
     4801 attgatgggg caaggccgca ggccgcgcag ttggagccgg tgggtatgtg gtcgaaggct
     4861 gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc tgtccatcag
     4921 cttgtccagc agggttgtcc acgggccgag cgaagcgagc cagccggtgg ccgctcgcgg
     4981 ccatcgtcca catatccacg ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc
     5041 ggagagcaag cccgtagggc gccgcagccg ccgtaggcgg tcacgacttt gcgaagcaaa
     5101 gtctagtgag tatactcaag cattgagtgg cccgccggag gcaccgcctt gcgctgcccc
     5161 cgtcgagccg gttggacacc aaaagggagg ggcaggcatg gcggcatacg cgatcatgcg
     5221 atgcaagaag ctggcgaaaa tgggcaacgt ggcggccagt ctcaagcacg cctaccgcga
     5281 gcgcgagacg cccaacgctg acgccagcag gacgccagag aacgagcact gggcggccag
     5341 cagcaccgat gaagcgatgg gccgactgcg cgagttgctg ccagagaagc ggcgcaagga
     5401 cgctgtgttg gcggtcgagt acgtcatgac ggccagcccg gaatggtgga agtcggccag
     5461 ccaagaacag caggcggcgt tcttcgagaa ggcgcacaag tggctggcgg acaagtacgg
     5521 ggcggatcgc atcgtgacgg ccagcatcca ccgtgacgaa accagcccgc acatgaccgc
     5581 gttcgtggtg ccgctgacgc aggacggcag gctgtcggcc aaggagttca tcggcaacaa
     5641 agcgcagatg acccgcgacc agaccacgtt tgcggccgct gtggccgatc tagggctgca
     5701 acggggcatc gagggcagca aggcacgtca cacgcgcatt caggcgttct acgaggccct
     5761 ggagcggcca ccagtgggcc acgtcaccat cagcccgcaa gcggtcgagc cacgcgccta
     5821 tgcaccgcag ggattggccg aaaagctggg aatctcaaag cgcgttgaga cgccggaagc
     5881 cgtggccgac cggctgacaa aagcggttcg gcaggggtat gagcctgccc tacaggccgc
     5941 cgcaggagcg cgtgagatgc gcaagaaggc cgatcaagcc caagagacgg cccgag
//
