LOCUS       .                       7626 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      complement(182..770)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(944..1801)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(1802..1906)
                     /label="AmpR promoter"
     rep_origin      complement(1932..2387)
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2899..3176
                     /label="chicken β-actin promoter"
     CDS             3597..4295
                     /label="TagBFP"
                     /note="monomeric blue fluorescent protein"
     misc_RNA        4394..4412
                     /label="MS2 stem loop"
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     CDS             4827..4880
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5418..5438
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             5808..6509
                     /label="TEV protease"
                     /note="tobacco etch virus protease (Kapust et al., 2001)"
     misc_feature    6528..7116
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    complement(7159..7280)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2761 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2821 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2881 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     2941 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     3001 gggcgggggg gggggggggg cgcgcgccgg gcggggcggg gcggggcgag gggcggggcg
     3061 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     3121 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     3181 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     3241 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     3301 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     3361 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     3421 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3481 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3541 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc gccaccatga
     3601 gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc
     3661 atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
     3721 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct
     3781 tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc
     3841 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
     3901 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
     3961 gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
     4021 ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac tacatggccc
     4081 tgaagctcgt gggcgggagc catctgatcg caaacgccaa gaccacatat agatccaaga
     4141 aacccgctaa taacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa
     4201 tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact
     4261 gcgacctccc tagcaaactg gggcacaagc tcaatactag tgctactaac ttcagcctgc
     4321 tgaagcaggc tggagacgtg gaggagaacc ctggacctga attcattggt aacagcggta
     4381 aaaagggact cagacatgag gatcacccat gtgctgtaca gacttggccc ccaataccaa
     4441 agggtggagg aacaccccac ccttccatat acctgaaagc caaacagtag gggaaagccc
     4501 tacgaaccac tgaaacagaa gcaccatcag ggcttctggg cactagtaaa ctgagccagg
     4561 agaaagtgcg tggagcatca gcccacgcat gcggcccact cccatacgaa ttttccgaat
     4621 tcactccacc aggtagtggc cgccgtcctt cagcttcagg gcctggtgga tctcgccctt
     4681 cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tagtcttctt
     4741 ctgcattacg gggccgtcgg gggggaagtt ggtgccgcgc atcttcacct tgtagatcag
     4801 cgtgccgtcc tgcagggggc gcgccagagg gcagaggaag tcttctaaca tgcggtgacg
     4861 tggaggagaa tcccggccct ttaattaacg ccaccatgga gaacactgaa aactcagtgg
     4921 attcaaaatc cattaaaaat ttggaaccaa agatcataca tggaagcgaa tcaatggact
     4981 ctggaatatc cctggacaac agttataaaa tggattatcc tgagatgggt ttatgtataa
     5041 taattaataa taagaatttt cataaaagca ctggaatgac atctcggtct ggtacagatg
     5101 tcgatgcagc aaacctcagg gaaacattca gaaacttgaa atatgaagtc aggaataaaa
     5161 atgatcttac acgtgaagaa attgtggaat tgatgcgtga tgtttctaaa gaagatcaca
     5221 gcaaaaggag cagttttgtt tgtgtgcttc tgagccatgg tgaagaagga ataatttttg
     5281 gaacaaatgg acctgttgac ctgaaaaaaa taacaaactt tttcagaggg gatcgttgta
     5341 gaagtctaac tggaaaaccc aaacttttca ttattcaggc ctgccgtggt acagaactgg
     5401 actgtggcat tgagacagaa aatctctact tccagagtgg tgttgatgat gacatggcgt
     5461 gtcataaaat accagtggag gccgacttct tgtatgcata ctccacagca cctggttatt
     5521 attcttggcg aaattcaaag gatggctcct ggttcatcca gtcgctttgt gccatgctga
     5581 aacagtatgc cgacaagctt gaatttatgc acattcttac ccgggttaac cgaaaggtgg
     5641 caacagaatt tgagtccttt tcctttgacg ctacttttca tgcaaagaaa cagattccat
     5701 gtattgtttc catgctcaca aaagaactct atttttatca ctccggaagc ggagagggca
     5761 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctggc gagagccttt
     5821 tcaagggccc gagggactac aacccgatct ccagcaccat ctgtcacctg accaacgaga
     5881 gcgacggtca caccactagt ctgtacggca tcggcttcgg ccccttcatc atcaccaaca
     5941 agcatctgtt caggaggaat aacggcacac tgctggtgca aagcctgcac ggcgtgttca
     6001 aagtgaagaa cacaaccacc ctgcaacagc acctgatcga cggcagggac atgattatca
     6061 tcaggatgcc caaggacttc cccccctttc cccagaaact gaagttcagg gagccacaaa
     6121 gggaggagcg aatctgcctg gtgaccacca acttccagac caagtccatg agcagcatgg
     6181 tctctgatac cagctgcacc ttccccagca gcgacggcat cttctggaag cactggattc
     6241 agacgaagga tggccaatgc ggcagcccat tggtgagcac tagggacggc ttcatcgtgg
     6301 gcatccacag cgccagcaat tttaccaata ccaacaacta cttcacgagc gtgccgaaaa
     6361 acttcatgga gctgttgacc aatcaagagg cgcagcagtg ggtgagcggc tggaggctga
     6421 acgccgacag cgttctttgg ggcggacata aggtgttcat ggtcaagccc gaggaaccct
     6481 tccagcccgt taaggaagcc actcagcttt gataaaagct tatcgataat caacctctgg
     6541 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     6601 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6661 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6721 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6781 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6841 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6901 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6961 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     7021 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     7081 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc
     7141 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     7201 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     7261 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     7321 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     7381 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     7441 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     7501 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     7561 cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7621 ggcgct
//
