LOCUS       V027827                 6140 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027827
VERSION     V027827
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..2427
                     /label="TFEB(NM_007162)"
                     /note="TFEB(NM_007162)"
                     /gene="TFEB"
     polyA_signal    2481..3103
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3132..3461
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(3500..3518)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3532..3548)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3556..3572
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /label="lac operator"
     promoter        complement(3580..3610)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3625..3646
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(3934..4522)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4693..5553)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(5554..5658)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5685..6113
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tggcgtcacg catagggttg
     1021 cgcatgcagc tcatgcggga gcaggcgcag caggaggagc agcgggagcg catgcagcaa
     1081 caggctgtca tgcattacat gcagcagcag cagcagcagc aacagcagca gctcggaggg
     1141 ccgcccaccc cggccatcaa tacccccgtc cacttccagt cgccaccacc tgtgcctggg
     1201 gaggtgttga aggtgcagtc ctacctggag aatcccacat cctaccatct gcagcagtcg
     1261 cagcatcaga aggtgcggga gtacctgtcc gagacctatg ggaacaagtt tgctgcccac
     1321 atcagcccag cccagggctc tccgaaaccc ccaccagccg cctccccagg ggtgcgagct
     1381 ggacacgtgc tgtcctcctc cgctggcaac agtgctccca atagccccat ggccatgctg
     1441 cacattggct ccaaccctga gagggagttg gatgatgtca ttgacaacat tatgcgtctg
     1501 gacgatgtcc ttggctacat caatcctgaa atgcagatgc ccaacacgct acccctgtcc
     1561 agcagccacc tgaatgtgta cagcagcgac ccccaggtca cagcctccct ggtgggcgtc
     1621 accagcagct cctgccctgc ggacctgacc cagaagcgag agctcacaga tgctgagagc
     1681 agggccctgg ccaaggagcg gcagaagaaa gacaatcaca acttaattga aaggagacga
     1741 aggttcaaca tcaatgaccg catcaaggag ttgggaatgc tgatccccaa ggccaatgac
     1801 ctggacgtgc gctggaacaa gggcaccatc ctcaaggcct ctgtggatta catccggagg
     1861 atgcagaagg acctgcaaaa gtccagggag ctggagaacc actctcgccg cctggagatg
     1921 accaacaagc agctctggct ccgtatccag gagctggaga tgcaggctcg agtgcacggc
     1981 ctccctacca cctccccgtc cggcatgaac atggctgagc tggcccagca ggtggtgaag
     2041 caggagctgc ctagcgaaga gggcccaggg gaggccctga tgctgggggc tgaggtccct
     2101 gaccctgagc cactgccagc tctgcccccg caagccccgc tgcccctgcc cacccagcca
     2161 ccatccccat tccatcacct ggacttcagc cacagcctga gctttggggg cagggaggac
     2221 gagggtcccc cgggctaccc cgaacccctg gcgccggggc atggctcccc attccccagc
     2281 ctgtccaaga aggatctgga cctcatgctc ctggacgact cactgctacc gctggcctct
     2341 gatccacttc tgtccaccat gtcccccgag gcctccaagg ccagcagccg ccggagcagc
     2401 ttcagcatgg aggagggcga tgtgctgtga gaattcatcg atagatctga tatcggtacc
     2461 agtcgactct agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     2521 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     2581 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     2641 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     2701 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     2761 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     2821 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     2881 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     2941 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     3001 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     3061 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     3121 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3301 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3361 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3421 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     3481 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     3541 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3601 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3661 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3721 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3781 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3841 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3901 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3961 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4021 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4081 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4141 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4201 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4261 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4321 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4381 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4441 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4501 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4561 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4621 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4681 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4801 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4861 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4981 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5221 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5401 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5461 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5521 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5581 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5641 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     5701 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5761 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5821 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5881 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5941 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6001 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6061 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6121 aatattaacg cttacaattt
//
