LOCUS       62056_3710        5930 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 5930)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5930
                     /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        1829..1857
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             1904..2620
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     2626..2642
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2676..2692)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2725..2743)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2753..2769)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3479..4138)
                     /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(4139..4908)
                     /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 tgggtacctt gacaattaat catcggctcg tataatgtgt
     1861 ggaaaagcgt gacgactcat gtttcacaca ggaaacagta ttcatggtga gcaagggcga
     1921 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1981 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2041 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2101 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2161 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2221 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2281 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2341 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2401 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2461 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2521 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2581 cgccgccggg atcactctcg gcatggacga gctgtacaag taactcgagg tcgacggtat
     2641 cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc
     2701 caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     2761 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2821 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2881 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt
     2941 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgact gcgatgagtg
     3001 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggtgctac
     3061 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg
     3121 tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt
     3181 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     3241 aggctctccc cgtggaggta ataattgacg atatgatcat ttattctgcc tcccagagcc
     3301 tgataaaaac ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     3361 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgaggcg
     3421 gctacagccg atagtctgga acagcgcact tacgggttgc tgcgcaaccc aagtgctacc
     3481 ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc gccatcgtcc agaaaacacg
     3541 gctcatcggg catcggcagg cgctgctgcc cgcgccgttc ccattcctcc gtttcggtca
     3601 aggctggcag gtctggttcc atgcccggaa tgccgggctg gctgggcggc tcctcgccgg
     3661 ggccggtcgg tagttgctgc tcgcccggat acagggtcgg gatgcggcgc aggtcgccat
     3721 gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca ctgaacaccg
     3781 acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc acgtaggccg
     3841 acacggtgcc ggggccgttg agcttcacga cggagatcca gcgctcggcc accaagtcct
     3901 tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt gtcttctggc
     3961 tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc agcattgccg
     4021 ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt tccgtttgca
     4081 cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc gtggccatgc
     4141 ttatctccat gcggtagggt gccgcacggt tgcggcacca tgcgcaatca gctgcaactt
     4201 ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta cagattttct
     4261 ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg tacccccctt
     4321 ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt ctttggtgcc
     4381 caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc cctccggcaa
     4441 aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg
     4501 cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg
     4561 cttcgccttc gactgccccc actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca
     4621 agccgctgcg cggtcgctgc gcgagccttg acccgccttc cacttggtgt ccaaccggca
     4681 agcgaagcgc gcaggccgca ggccggaggc ttttccccag agaaaattaa aaaaattgat
     4741 ggggcaaggc cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa ggctgggtag
     4801 ccggtgggca atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc
     4861 cagcagggtt gtccacgggc cgagcgaagc gagccagccg gtggccgctc gcggccatcg
     4921 tccacatatc cacgggctgg caagggagcg cagcgaccgc gcagggcgaa gcccggagag
     4981 caagcccgta gggcgccgca gccgccgtag gcggtcacga ctttgcgaag caaagtctag
     5041 tgagtatact caagcattga gtggcccgcc ggaggcaccg ccttgcgctg cccccgtcga
     5101 gccggttgga caccaaaagg gaggggcagg catggcggca tacgcgatca tgcgatgcaa
     5161 gaagctggcg aaaatgggca acgtggcggc cagtctcaag cacgcctacc gcgagcgcga
     5221 gacgcccaac gctgacgcca gcaggacgcc agagaacgag cactgggcgg ccagcagcac
     5281 cgatgaagcg atgggccgac tgcgcgagtt gctgccagag aagcggcgca aggacgctgt
     5341 gttggcggtc gagtacgtca tgacggccag cccggaatgg tggaagtcgg ccagccaaga
     5401 acagcaggcg gcgttcttcg agaaggcgca caagtggctg gcggacaagt acggggcgga
     5461 tcgcatcgtg acggccagca tccaccgtga cgaaaccagc ccgcacatga ccgcgttcgt
     5521 ggtgccgctg acgcaggacg gcaggctgtc ggccaaggag ttcatcggca acaaagcgca
     5581 gatgacccgc gaccagacca cgtttgcggc cgctgtggcc gatctagggc tgcaacgggg
     5641 catcgagggc agcaaggcac gtcacacgcg cattcaggcg ttctacgagg ccctggagcg
     5701 gccaccagtg ggccacgtca ccatcagccc gcaagcggtc gagccacgcg cctatgcacc
     5761 gcagggattg gccgaaaagc tgggaatctc aaagcgcgtt gagacgccgg aagccgtggc
     5821 cgaccggctg acaaaagcgg ttcggcaggg gtatgagcct gccctacagg ccgccgcagg
     5881 agcgcgtgag atgcgcaaga aggccgatca agcccaagag acggcccgag
//
