LOCUS       V021402                 6341 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021402
VERSION     V021402
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             935..958
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             962..2131
                     /label="Pdha1(NM_008810)"
                     /note="Pdha1(NM_008810)"
                     /gene="Pdha1"
     CDS             2153..2671
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNG
                     IKANFKIRHNIE"
                     /label="VN173"
     polyA_signal    2682..3304
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3333..3662
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(3701..3719)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3733..3749)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3757..3773
                     /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(3781..3811)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    3826..3847
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4135..4723)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(4894..5754)
                     /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(5755..5859)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5886..6314
                     /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 attgatctac catggactac aaagacgatg acgacaagct
      961 tatgaggaag atgcttgccg ctgtatcccg cgtgttggca ggctctgcgc agaagccggc
     1021 aagccgagtg ctggttgctt cccgtaattt tgcaaatgat gctacatttg agattaagaa
     1081 atgtgacctt catcggctag aagagggccc cccagtcacc acagtgctca ccagagagga
     1141 tgggctcaag tactacagga tgatgcagac tgtgcgccgg atggagctaa aggcggatca
     1201 gctgtataag cagaaaatca ttcgtggttt ctgtcacttg tgtgatggtc aggaagcctg
     1261 ctgcgtgggc ctggaggctg gcataaaccc tacggaccac ctcatcactg cctatcgagc
     1321 acatggcttc accttcactc ggggcctgcc tgtgcgagca attcttgcag agctaacagg
     1381 acgaagagga ggttgtgcta aagggaaagg cgggtcaatg cacatgtacg ccaagaactt
     1441 ctatggaggc aacggcatcg ttggagctca ggtgcccctg ggagcaggaa ttgccctggc
     1501 ctgcaagtac aatggaaaag atgaggtctg tttgacatta tacggcgatg gtgctgctaa
     1561 tcagggtcag atctttgaag cttacaatat ggcagcactg tggaaattac cttgcatttt
     1621 tatctgtgag aacaaccgct atggcatggg gacgtctgtt gagagagcag cagccagcac
     1681 ggactactac aaaagaggag attttattcc tggactcagg gtagatggaa tggatatctt
     1741 gtgcgtccga gaggcaacaa agtttgcggc tgcctattgc aggtctggta aggggcccat
     1801 cctgatggag ctccagactt accgctacca tggacacagc atgagtgacc ctggagtaag
     1861 ctaccgcact cgagaagaaa tccaggaagt aagaagtaag agtgacccta ttatgcttct
     1921 caaggataga atggtgaaca gcaatcttgc aagtgttgaa gaattaaagg agattgatgt
     1981 ggaagtgagg aaagaaatcg aggatgctgc ccagtttgcc acggctgatc ctgagccccc
     2041 gttggaggaa ctaggctatc acatctacag cagtgatcct ccctttgaag tgcgtggtgc
     2101 caaccagtgg atcaagttta agtcagtcag ttctagaaga tccatcgcca ccatggtgag
     2161 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2221 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2281 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2341 caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     2401 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2461 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2521 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2581 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     2641 ggccaacttc aagatccgcc acaacatcga gtagggatcc cgggtggcat ccctgtgacc
     2701 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2761 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2821 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2881 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2941 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3001 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3061 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3121 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     3181 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     3241 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     3301 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3421 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3481 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3541 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3601 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3661 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     3721 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3781 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3841 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5881 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5941 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6001 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6061 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6121 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6181 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6241 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6301 atttaacgcg aattttaaca aaatattaac gcttacaatt t
//
