LOCUS       V047607                 6709 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047607
VERSION     V047607
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             964..2199
                     /label="Pdk4(NM_053551)"
                     /note="Pdk4(NM_053551)"
                     /gene="Pdk4"
     CDS             2206..2271
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2309..2533
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2579..3007
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3021..3350
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3417..4211
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4385..4506
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4555..4571)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4579..4595
                     /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(4603..4633)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4648..4669
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4957..5542)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5713..6573)
                     /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(6574..6678)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgaagg cagcccgttt cgtgatgcgc agcgccagct cgctgggcaa cgccggcctg
     1021 gtccccaggg aggtcgagct gttctcccgc tacagcccgt ccccgctgtc catgaagcag
     1081 ctgctggact tcggttcaga aaatgcctgt gagagaactt ctttttcttt tcttcggcaa
     1141 gaattgcccg tcagacttgc caacatcctg aaggagattg acatcctccc tgaacgctta
     1201 gtgaacaccc cttccgtcca gctggtgaag agctggtaca tccagagcct gatggattta
     1261 gtggagttcc acgaaaagag cccagaagac cagaaagctc tatcagactt tgtagacacg
     1321 ctggtcaaag ttcgaaacag acatcataat gtggtcccta cgatggcaca agggatcctg
     1381 gagtacaaag acaactgcac cgttgacccc gttaccaatc aaaatcttca gtattttttg
     1441 gaccggtttt atatgaaccg catttctact cggatgctca tgaaccagca catcctcata
     1501 ttcagtgact caaagacagg aaacccaagc cacattggga gtatcgaccc caactgcgat
     1561 gtggtagcag tagtcgaaga tgcctttgag tgtgcaaaga tgctctgcga tcagtattat
     1621 ctaacgtcgc cagaattaaa gctcacacaa gtcaatggaa aatttccagg ccaaccaatc
     1681 cacatcgtgt acgttccttc acaccttcac cacatgctct ttgaactatt caagaatgcc
     1741 atgagggcca cggtcgagca tcaagaaaac cgcccttccc tgacaccagt cgaggccacc
     1801 gtcgtcttgg gaaaagaaga tcttacaatc aagatttctg accgaggagg cggtgttcct
     1861 ctgaggatta ctgaccgcct ctttagttac acatactcca ctgctccaac gcctgtgatg
     1921 gacaattcac ggaatgcccc tttggctggt tttggttacg gcttgccgat ttcccgtctc
     1981 tacgccaagt attttcaagg agatctgaat ctctattcca tgtcaggcta tgggacagat
     2041 gccatcatct acttaaaggc tttatcatct gagtctatag aaaaactccc cgtctttaac
     2101 aagtcagcct tcaaacatta tcagatgagt tctgaggctg atgactggtg tatcccaagt
     2161 aaggaaccaa agaatctgtc gaaggagaaa ctggcagtgc tcgaggacta caaagaccat
     2221 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg
     2281 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2341 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2401 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2461 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2521 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2581 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2641 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2701 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2761 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2821 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2881 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2941 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3001 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3181 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3241 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3301 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3361 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3421 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3481 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3541 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3601 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3661 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3721 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3781 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3841 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3901 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3961 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4021 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4081 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4141 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4201 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4261 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4321 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4381 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4441 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4501 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4561 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4621 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4741 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4801 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4861 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5461 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5881 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6001 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6241 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6301 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6421 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6541 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6661 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
