LOCUS       .                       7617 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"
     promoter        1382..1731
                     /label="TRE3GV promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On® 3G, optimized for
                     retroviral and lentiviral vectors"
     CDS             1744..2451
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             2476..3540
                     /label="lacI"
                     /note="lac repressor"
     CDS             3556..3576
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     promoter        3702..3913
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             3926..4194
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4227..4622
                     /label="BSD"
                     /note="blasticidin S deaminase"
     polyA_signal    4678..4799
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5596..5612)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5620..5636)
                     /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(5644..5674)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5689..5710)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5998..6586)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6760..7617)
                     /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 tagtcccttt cgtcttcact
     1381 cgagtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     1441 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     1501 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     1561 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     1621 gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag
     1681 tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatact tggatccgcc
     1741 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1801 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1861 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1921 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1981 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2041 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2101 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2161 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2221 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2281 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2341 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2401 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggactc
     2461 agatctcgag ggatggtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     2521 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     2581 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     2641 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     2701 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     2761 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     2821 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     2881 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     2941 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     3001 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     3061 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     3121 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     3181 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     3241 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     3301 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     3361 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     3421 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     3481 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3541 agcagcctga ggcctcctaa gaagaagagg aaggttggta ccgcgggcgc tggtggcctg
     3601 aatgacatct ttgaggccca gaagatcgag tggcatgagg gatccaccgg atctagataa
     3661 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3721 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     3781 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3841 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     3901 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     3961 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4021 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4081 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     4141 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     4201 gtgaccggcg cctacgctag gccaccatgg ccaagccttt gtctcaagaa gaatccaccc
     4261 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     4321 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     4381 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     4441 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     4501 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     4561 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4621 gctaagaatt ctacctacgc tatgtggata cgctgcttta atgcctttgt atcagttaac
     4681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4801 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     4861 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     4921 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     4981 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     5041 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     5101 tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg ctcagtcatc
     5161 gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg
     5221 ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac gttgagcgaa
     5281 aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt aacggtgaac
     5341 tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca cagttccgga
     5401 tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg aactgccgtg
     5461 ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc gaggacgggt
     5521 atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc ggcgggcgcg
     5581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6421 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcat
//
