LOCUS       62056_12545        3628 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 3628)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             579..601
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             620..637
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             647..1363
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1407..1423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1431..1447)
                     /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(1455..1485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1500..1521)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1809..2397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2571..3428)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3429..3533)
                     /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 ccagtgccaa gcttgcatgc ctgcaggtcg
      421 acgacccgca cgaccagcgt tagcatgctc agtaagttga gtgcatcagg ctcagctctg
      481 aattgacagc acaccgccgt cgaggcaagc ttgagcgggg tgcactcatc atagtgcagg
      541 aaagaagctc tacatattca ggaggattca ccggatcctt tgtttaactt taagaaggag
      601 atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcatgg tgagcaaggg
      661 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      721 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      781 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      841 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      901 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      961 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1021 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1081 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1141 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1201 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1261 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1321 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatttt tttataccgg
     1381 gtaccgagct cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1441 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1501 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1561 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1621 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1681 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1741 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1801 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1861 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1921 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1981 tcgggaagcg tggcgctttc tcaaagctca cgctgtaggt atctcagttc ggtgtaggtc
     2041 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2101 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2161 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2221 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2281 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2341 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2401 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2461 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2521 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2581 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2641 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2701 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2761 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2821 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2881 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2941 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3001 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3061 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3121 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3181 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3241 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3301 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3361 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3421 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3481 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3541 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3601 aataggcgta tcacgaggcc ctttcgtc
//
