LOCUS       .                       3894 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     terminator      412..483
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      499..526
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     RBS             602..624
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             643..660
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             670..687
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             733..1449
                     /label="EGFP"
                     /note="the original enhanced GFP (Yang et al., 1996)"
     CDS             1474..1491
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      1558..1605
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     primer_bind     complement(1673..1689)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1697..1713)
                     /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-β-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1721..1751)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1766..1787)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2075..2663)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2837..3694)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(3695..3799)
                     /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 cagccaggca tcaaataaaa
      421 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      481 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag
      541 taacaccgtg cgtgttgact attttacctc tggcggtgat aatggttgct ctagaaataa
      601 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
      661 agcagcggcc tggtgccgcg cggcagccat atggctagca tgactggtgg acagcaaatg
      721 ggtcggatcc atatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      781 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      841 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      901 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      961 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1021 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1081 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1141 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1201 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1261 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1321 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1381 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1441 ctgtacaagt gaaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg
     1501 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1561 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaagg
     1621 atcccgggcc cgtcgactgc agaggcctgc atgcaagctt ggcgtaatca tggtcatagc
     1681 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     1741 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1861 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1921 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1981 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2041 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2101 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2161 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2221 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2281 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2341 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2401 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2461 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2521 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2581 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2641 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2701 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2761 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2821 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2881 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2941 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3001 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3061 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3121 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3181 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3241 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3301 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3361 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3421 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3481 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3541 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3601 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3661 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3721 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3781 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3841 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
