LOCUS       pCIRCNpu                3817 bp    DNA     circular UNA 07-JAN-2016
DEFINITION  Vector for SICLOPPS-mediated protein circularization using the
            Nostoc punctiforme DnaE intein.
ACCESSION   urn.local...a-4oqv6v0 (modified)
VERSION     urn.local...a-4oqv6v0
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          1..3817
                     /organism="synthetic construct pCIRCNpu"
                     /mol_type="other DNA"
     regulatory      1..19
                     /regulatory_class="promoter"
                     /gene="T7 promoter"
     misc_feature    19..46
                     /gene="lac operator"
     misc_feature    47..68
                     /gene="BioBrick RFC[10] prefix"
     regulatory      80..96
                     /regulatory_class="RBS"
                     /gene="T7 RBS"
     mat_peptide     104..211
                     /product="Npu DnaE(C)"
     misc_feature    104..106
                     /note="Start"
     misc_feature    219..1287
                     /gene="BBa_J04450"
     misc_feature    219..418
                     /gene="BBa_R0010"
     misc_feature    307..344
                     /gene="CAP binding site"
     regulatory      355..360
                     /regulatory_class="minus_35_sigmal"
                     /note="-35 signal"
     regulatory      379..384
                     /regulatory_class="minus_10_signal"
                     /note="-10 signal"
     misc_feature    388..418
                     /gene="lac operator"
     regulatory      427..438
                     /regulatory_class="RBS"
                     /gene="BBa_B0034"
     misc_feature    431..434
                     /note="conserved"
     CDS             445..1119
                     /gene="mRFP1"
     misc_feature    445..1125
                     /gene="BBa_E1010"
     misc_feature    1126..1150
                     /note="barcode"
     misc_feature    1159..1238
                     /gene="BBa_B0010"
     regulatory      1170..1213
                     /regulatory_class="terminator"
                     /gene="stem loop"
     misc_feature    1247..1287
                     /gene="BBa_B0012"
     regulatory      1254..1273
                     /regulatory_class="terminator"
                     /gene="T7 TE"
     regulatory      1274..1287
                     /regulatory_class="polyA_signal_sequence"
                     /gene="polyA signal"
     misc_feature    1280
                     /note="stop"
     mat_peptide     1295..1600
                     /gene="Npu DnaE(N)"
     misc_feature    1631..1651
                     /gene="BioBrick RFC[10] suffix"
     regulatory      1652..1723
                     /regulatory_class="terminator"
                     /gene="E. coli his operon terminator"
     stem_loop       1660..1694
     primer_bind     complement(1785..1804)
                     /gene="VR"
     rep_origin      complement(1891..2505)
                     /gene="rep (pMB1)"
     CDS             complement(2727..3542)
                     /gene="Kanamycin resistance"
     misc_feature    3510
                     /note="Silent Mut: G->C mutation to remove XhoI site"
     misc            complement(3647..3690)
                     /gene="repeat region"
     primer_bind     3695..3714
                     /gene="Verification forward (VF2)"
     regulatory      complement(3773..3805)
                     /regulatory_class="terminator"
                     /gene="terminator"
     stem_loop       complement(3786..3805)
ORIGIN      
        1 taatacgact cactataggg gaattgtgag cggataacaa ttccccgaat tcgcggccgc
       61 ttctagagaa taattttgtt taactttaag aaggagatac tagatgatca aaatagccac
      121 acgtaaatat ttaggcaaac aaaatgtcta tgacattgga gttgagcgcg accataattt
      181 tgcactcaaa aatggcttca tagcttccaa cagagaccca atacgcaaac cgcctctccc
      241 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      301 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      361 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacata
      421 ctagagaaag aggagaaata ctagatggct tcctccgaag acgttatcaa agagttcatg
      481 cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa
      541 ggtgaaggtc gtccgtacga aggtacccag accgctaaac tgaaagttac caaaggtggt
      601 ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc agtacggttc caaagcttac
      661 gttaaacacc cggctgacat cccggactac ctgaaactgt ccttcccgga aggtttcaaa
      721 tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta ccgttaccca ggactcctcc
      781 ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac
      841 ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg tatgtacccg
      901 gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac
      961 tacgacgctg aagttaaaac cacctacatg gctaaaaaac cggttcagct gccgggtgct
     1021 tacaaaaccg acatcaaact ggacatcacc tcccacaacg aagactacac catcgttgaa
     1081 cagtacgaac gtgctgaagg tcgtcactcc accggtgctt aataacgctg atagtgctag
     1141 tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     1201 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     1261 cttcgggtgg gcctttctgc gtttataggt ctcatgctta agctatgaaa cggaaatatt
     1321 gacagtagaa tatggattat taccgattgg taaaattgta gaaaagcgca tcgaatgtac
     1381 tgtttatagc gttgataata atggaaatat ttatacacaa cctgtagcac aatggcacga
     1441 tcgcggagaa caagaggtgt ttgagtattg tttggaagat ggttcattga ttcgggcaac
     1501 aaaagaccat aagtttatga ctgttgatgg tcaaatgttg ccaattgatg aaatatttga
     1561 acgtgaattg gatttgatgc gggttgataa tttgccgaat taataaatcg gtgaaatgca
     1621 cgactgatag tactagtagc ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac
     1681 cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca
     1741 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1801 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1861 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac aggctccgcc
     1921 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1981 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2041 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2101 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2161 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2221 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2281 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2341 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2401 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2461 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2521 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2581 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2641 atgagtaaac ttggtctgac agctcgagtc ccgtcaagtc agcgtaatgc tctgccagtg
     2701 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     2761 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2821 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     2881 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     2941 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     3001 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3061 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3121 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3181 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3241 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3301 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3361 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     3421 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3481 atccatgttg gaatttaatc gcggcctgga gcaagacgtt tcccgttgaa tatggctcat
     3541 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     3601 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     3661 tcgaactttt gctgagttga aggatcagct cgagtgccac ctgacgtcta agaaaccatt
     3721 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
     3781 aaatccttag ctttcgctaa ggatgatttc tggaatt
//
