LOCUS       V047425                 6625 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047425
VERSION     V047425
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             925..2376
                     /label="Krt14(NM_016958)"
                     /note="Krt14(NM_016958)"
                     /gene="Krt14"
     CDS             2389..2454
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2489..2713
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2759..3187
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3201..3530
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3542..4141
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4301..4422
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4471..4487)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4495..4511
                     /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(4519..4549)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4564..4585
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4873..5458)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5629..6489)
                     /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(6490..6594)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcgc caccatggcc acctgcagcc gccagttcac ctcctccagc
      961 tccatgaagg gctcttgtgg tatcggtggt ggctctagcc gcatgtcctc catcctggct
     1021 ggaggatcct gccgggctcc cagcacctac gggggcatgt cggttacctc ctctcgcttc
     1081 tcctctgggg gagcctgtgg gattggaggt ggctatggtg ggagcttcag cagcagcagt
     1141 tttggtggag gacttggtag tggatttggt ggtcgatttg atggatttgg tggtggtttt
     1201 ggtggtggtc ttggtggtgg ttttggtggt ggtcttggtg gtggtcttgg cggtggtatt
     1261 ggtgatgggc tcctggtggg cagtgagaaa gtgaccatgc agaacctcaa tgaccgcctg
     1321 gccacctacc tggacaaggt gcgtgccctg gaagaggcca acactgaact ggaggtgaag
     1381 attcgggact ggtaccagag gcagcggccc actgagatca aagactacag cccctacttc
     1441 aagaccatcg aggacctgaa gagcaagatc ctggcagcca ccgtggacaa tgccaatgtc
     1501 ctcctgcaga tcgacaatgc ccgcctggct gccgatgact tccggaccaa gtttgagacg
     1561 gagcagagcc tgcgcatgag cgtggaggcc gacatcaatg gcctgcgcag ggtgctggat
     1621 gagctgaccc tggccagagc cgacctggag atgcagattg agagcctcaa ggaggagctg
     1681 gcctacctga agaagaacca cgaggaggaa atggcctcca tgagaggcca ggtgggtgga
     1741 gacgtcaatg tggagatgga cgccgcccct ggtgtggacc tgagccgcat cctgaacgag
     1801 atgcgggatc agtacgagaa gatggcggag aagaaccgca aggatgctga ggaatggttc
     1861 ttcagcaaga cagaggagct gaaccgcgag gtggccacca acagcgagct ggtgcagagc
     1921 ggcaagagtg agatttctga gctccggcgc accatgcaga acctggagat cgagctgcag
     1981 tcccagctca gcatgaaagc atccctggag aataacctgg aggagaccaa aggccgttac
     2041 tgcatgcagc tggcccagat ccaggagatg atcggcagtg tggaggagca gctggctcag
     2101 ctgcgctgcg agatggagca gcagaaccag gagtacaaaa tcctgctgga tgtgaagaca
     2161 aggctggagc aggagatcgc cacctaccgc cgtctgctgg agggagagga cgcccacctt
     2221 tcatcttccc aattctcctc atcctctcaa ttctcctctg gctctcagtc atccagagat
     2281 gtgacctcca ccaaccgcca gatccgcacc aaggtcatgg atgtgcacga tggcaaggtg
     2341 gtctccaccc acgagcaggt cctgcgcacc aagaacctcg agaccggtga ctacaaagac
     2401 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagtgaccg
     2461 gtaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2521 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2581 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2641 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2701 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2761 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2821 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2881 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2941 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3001 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3061 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3121 gggattttgg ccatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3181 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3361 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3421 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3481 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3541 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3601 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3661 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3721 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3781 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3841 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3901 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3961 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4021 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4081 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4141 attcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4201 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4261 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4441 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4501 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4561 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4621 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4681 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4741 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4801 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4861 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4921 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4981 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5041 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5101 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5161 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5221 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5281 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5341 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5401 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6601 tccccgaaaa gtgccacctg acgtc
//
