LOCUS       V024287                 7668 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024287
VERSION     V024287
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2348
                     /label="Pkm(NM_001378866)"
                     /note="Pkm(NM_001378866)"
                     /gene="Pkm"
     CDS             2355..2420
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2444..3032
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3098..3146
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3348..3776
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3790..4119
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4186..4980
                     /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    5156..5277
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5326..5342)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5350..5366
                     /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(5374..5404)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5419..5440
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5728..6316)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6487..7347)
                     /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(7348..7452)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc gaagccacac agtgaagcag
      781 ggactgcctt cattcagacc cagcagctcc atgcagccat ggctgacacc ttcctggaac
      841 acatgtgccg cctggacatt gactctgccc ccatcacggc ccgcaacact ggcatcattt
      901 gtaccattgg gcctgcttcc cgatctgtgg agatgctgaa ggagatgatt aagtctggaa
      961 tgaatgtggc tcggctgaat ttctctcatg gaacccatga gtaccatgca gagaccatca
     1021 agaatgtccg tgaagccaca gaaagctttg catctgatcc cattctctac cgtcctgttg
     1081 cggtggctct ggatacaaag ggacctgaga tccggactgg actcatcaag ggcagcggca
     1141 ccgctgaggt ggagctgaag aagggagcca ctctgaagat caccctggac aacgcttaca
     1201 tggagaagtg tgacgagaac atcctgtggc tggactacaa gaacatctgc aaggtggtgg
     1261 aggtgggcag caagatctac gtggacgatg ggctcatctc actgcaggtg aaggagaaag
     1321 gcgctgactt cctggtgacg gaggtggaga atggtggctc cttgggcagc aagaagggcg
     1381 tgaacctgcc gggcgctgct gtggatctcc ccgctgtgtc ggaaaaggac atccaggacc
     1441 tgaagtttgg ggtggagcag gatgtggaca tggtgtttgc atctttcatc cgcaaggcag
     1501 ccgacgtgca tgaagtcagg aaggtgctgg gagagaaggg caagaacatc aagatcatca
     1561 gcaaaatcga gaaccatgaa ggcgtccgca ggtttgatga gatcttggag gccagtgatg
     1621 ggatcatggt ggctcgtggt gacctgggca ttgagattcc tgcagagaag gtcttcctgg
     1681 ctcagaagat gatgatcggg cgatgcaacc gagctgggaa gcctgtcatc tgtgccacac
     1741 agatgctgga gagcatgatc aagaagccac gccccacccg tgctgaaggc agtgatgtgg
     1801 ccaatgcagt cctggatgga gcagactgca tcatgctgtc tggagaaaca gccaaggggg
     1861 actaccctct ggaggctgtt cgcatgcagc acctgatagc tcgggaggct gaggcagcca
     1921 tgttccaccg tctgctgttt gaagagcttg tgcgagcctc cagtcactcc acagacctca
     1981 tggaggccat ggccatgggc agcgtggagg cctcttataa gtgtttagca gcagctttga
     2041 tagttctcac ggagtctggc aggagtgctc accaagtggc caggtaccgc cctcgggctc
     2101 ctatcattgc cgtgactcga aatccccaga ctgctcgcca ggcccatctg taccgtggca
     2161 tcttccctgt gctgtgtaag gatgccgtgc tgaatgcctg ggctgaggat gtcgaccttc
     2221 gtgtaaactt ggccatggat gttggcaagg cccgaggctt cttcaagaag ggagatgtgg
     2281 tcattgtgct gaccgggtgg cgccctggct ctggattcac caacaccatg cgtgtagtgc
     2341 ctgtacctgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2401 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2821 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3001 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3061 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3121 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3181 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3241 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3301 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3361 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3421 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3481 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3541 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3601 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3661 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3721 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3781 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3841 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3961 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4021 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4081 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4141 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4201 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4261 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4321 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4381 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4441 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4501 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4561 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4621 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4681 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4741 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4801 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4861 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4921 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4981 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5041 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5101 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5281 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5341 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5401 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5461 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5521 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7501 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7561 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7621 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
