LOCUS       .                       8259 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(1..105)
                     /label="AmpR promoter"
     rep_origin      complement(131..586)
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     intron          2100..3111
                     /label="chimeric intron"
                     /note="chimera between introns from chicken β-actin and
                     rabbit β-globin"
     CDS             3368..4009
                     /label="GFP(1-10)"
                     /note="fragment of superfolder GFP consisting of the first
                     10 β-strands"
     CDS             4069..4122
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4123..4719
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    4830..4962
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6238..6254)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6262..6278)
                     /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(6286..6316)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6331..6352)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6640..7228)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7402..8259)
                     /label="AmpR"
                     /note="β-lactamase"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     1501 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     1561 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     1621 actttccatt gacgtcaatg ggtggactat ttacggtaaa ctgcccactt ggcagtacat
     1681 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1741 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1801 ttagtcatcg ctattaccat gggtcgaggt gagccccacg ttctgcttca ctctccccat
     1861 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
     1921 gatgggggcg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     1981 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     2041 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     2101 gagtcgctgc gttgccttcg ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc
     2161 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     2221 gctgtaatta gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc
     2281 ttaaagggct ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg
     2341 tgtgtgtgcg tggggagcgc cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg
     2401 ggcgcggcgc ggggctttgt gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg
     2461 tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     2521 ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt aacccccccc tgcacccccc
     2581 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc
     2641 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     2701 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc cccggagcgc cggcggctgt
     2761 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     2821 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     2881 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     2941 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     3001 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3061 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3121 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattctgcag tcgacggtac
     3181 ccgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     3241 aagattctag agctagcgaa ttcgaattta aatcggatcc gccaccatgc tggccacccg
     3301 cgtgttcagc ctggtgggca agcgcgccat cagcaccagc gtgtgcgtgc gcgcccacaa
     3361 ggatccaatg tccaaaggag aagaactgtt taccggtgtt gtgccaattt tggttgaact
     3421 cgatggtgat gtcaacggac ataagttctc agtgagaggc gaaggagaag gtgacgccac
     3481 cattggaaaa ttgactctta aattcatctg tactactggt aaacttcctg taccatggcc
     3541 gactctcgta acaacgctta cgtacggagt tcagtgcttt tcgagatacc cagaccatat
     3601 gaaaagacat gactttttta agtcggctat gcctgaaggt tacgtgcaag aaagaacaat
     3661 ttcgttcaaa gatgatggaa aatataaaac tagagcagtt gttaaatttg aaggagatac
     3721 tttggttaac cgcattgaac tgaaaggaac agattttaaa gaagatggta atattcttgg
     3781 acacaaactc gaatacaatt ttaatagtca taacgtatac atcactgctg ataagcaaaa
     3841 gaacggaatt aaagcgaatt tcacagtacg ccataatgta gaagatggca gtgttcaact
     3901 tgccgaccat taccaacaaa acacccctat tggagacggt ccggtacttc ttcctgataa
     3961 tcactacctc tcaacacaaa cagtcctgag caaagatcca aatgaaaaag gaacatagcc
     4021 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga
     4081 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc
     4141 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4201 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4261 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4321 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4381 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4441 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4501 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4561 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4621 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4681 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac
     4741 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4801 tacgctgctt taatgccttt gtatcagtta acttgtttat tgcagcttat aatggttaca
     4861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattgactc aaatgatgtc
     4981 aattagtcta tcagaagcta tctggtctcc cttccggggg acaagacatc cctgtttaat
     5041 atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt
     5101 gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta
     5161 gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt tccccgtatc
     5221 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt
     5281 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag
     5341 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt
     5401 aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat
     5461 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     5521 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     5581 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     5641 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     5701 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc
     5761 atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc
     5821 tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag
     5881 agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga
     5941 tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct
     6001 gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca
     6061 tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg
     6121 acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc
     6181 agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc
     6241 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6301 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7081 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7261 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7321 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7381 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7501 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7561 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7921 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8221 aaagggaata agggcgacac ggaaatgttg aatactcat
//
