LOCUS       62056_22685        4025 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 4025)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             77..793
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             821..838
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             845..871
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     terminator      911..958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1132..1920)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1921..2012)
                     /label=AmpR promoter
     rep_origin      2021..2609
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(2769..2790)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2806..3885)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3886..3963)
                     /label=lacI promoter
     promoter        4009..4025
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc ggtaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      121 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      181 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      241 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      301 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      361 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      421 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      481 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      541 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      601 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      661 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      721 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      781 cgagctgtac aagtaagaat tcgcggccgc agagctcgct ctggtgccac gcggtagttc
      841 cgctcatcac caccatcatc accatcacca ccactaatta acctaggctg ctgccaccgc
      901 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      961 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
     1021 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc
     1081 atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt atttgccgac
     1141 taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga
     1201 ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg
     1261 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     1321 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     1381 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     1441 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     1501 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     1561 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     1621 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     1681 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     1741 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     1801 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     1861 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     1921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1981 catatttgaa tgtatttaga aaaataaaca aatagctagc ttgagatcct ttttttctgc
     2041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2161 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2581 gctcgtcagg ggggcggagc ctatggaaat ctagatttca gtgcaattta tctcttcaaa
     2641 tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc
     2701 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     2761 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     2821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2881 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     2941 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     3001 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     3061 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     3121 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     3181 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     3241 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     3301 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     3361 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     3421 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     3481 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     3541 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     3601 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     3661 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     3721 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     3781 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     3841 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     3901 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     3961 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta atacgactca
     4021 ctata
//
