LOCUS       62056_22315        4478 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 4478)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(435..512)
                     /label=lacI promoter
     promoter        558..586
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             600..1298
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1329..2045
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      2115..2162
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2257..2273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2281..2297)
                     /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)."
     promoter        complement(2305..2335)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2350..2371)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3421..4278)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4279..4383)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
      481 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
      541 ctcctgcatt aggaaatttg acaattaatc atcggctcgt ataatgtgtg gaatgtctca
      601 tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtggaca
      661 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
      721 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
      781 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
      841 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
      901 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
      961 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     1021 aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
     1081 ccctgaagct cgtgggcggg agccatctga tcgcaaacgc caagaccaca tatagatcca
     1141 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
     1201 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
     1261 actgcgacct ccctagcaaa ctggggcaca agcttaatgg cggtggagga agcggcggag
     1321 gtggatctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1381 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1441 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1501 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1561 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1621 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1681 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1741 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1801 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1861 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1921 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1981 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2041 acaagtagtc caccggcggc atggacgagc tgtacaagta attaacctag gctgctgcca
     2101 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2161 tgctgaaacc tcaggcattt gagaagcaca cggtcacaca tcggatcccg ggcccgtcga
     2221 ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2281 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2341 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4381 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4441 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
