LOCUS       .                       4256 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"
     promoter        618..636
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             668..690
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             709..726
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             736..747
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             766..1473
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      1572..1619
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        complement(1985..2013)
                     /label="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     primer_bind     complement(2035..2051)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(2059..2075)
                     /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(2083..2113)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2128..2149)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2437..3025)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3199..4056)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(4057..4161)
                     /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 aaggagatgg
      421 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      481 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      541 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
      601 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
      661 aaataatttt gtttaacttt aagaaggaga tataccatgt ctggttctca tcatcatcat
      721 catcatagca gcggcatcga aggccgcggc cgcttaatta aaaccatggt gagcaagggc
      781 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
      841 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      901 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
      961 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     1021 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1081 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     1141 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     1201 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     1261 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     1321 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     1381 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     1441 cactccaccg gcggcatgga cgagctgtac aagtagctcg agcgagctct gcagcccggg
     1501 atccgtaaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1561 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1621 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
     1681 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     1741 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     1801 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     1861 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     1921 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     1981 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcgtaat catggtcata
     2041 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2101 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2161 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2221 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2281 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2341 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2401 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3481 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3781 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4201 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
