LOCUS       V024491                 7109 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024491
VERSION     V024491
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             989..2581
                     /label="PKM(NM_002654)"
                     /note="PKM(NM_002654)"
                     /gene="PKM"
     CDS             2594..2623
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2639..2656
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2706..2930
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2976..3404
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3418..3747
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3814..4608
                     /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    4782..4903
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4952..4968)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4976..4992
                     /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(5000..5030)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5045..5066
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5354..5942)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6113..6973)
                     /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(6974..7078)
                     /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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtggat ccgccaccat gtcgaagccc catagtgaag ccgggactgc
     1021 cttcattcag acccagcagc tgcacgcagc catggctgac acattcctgg agcacatgtg
     1081 ccgcctggac attgattcac cacccatcac agcccggaac actggcatca tctgtaccat
     1141 tggcccagct tcccgatcag tggagacgtt gaaggagatg attaagtctg gaatgaatgt
     1201 ggctcgtctg aacttctctc atggaactca tgagtaccat gcggagacca tcaagaatgt
     1261 gcgcacagcc acggaaagct ttgcttctga ccccatcctc taccggcccg ttgctgtggc
     1321 tctagacact aaaggacctg agatccgaac tgggctcatc aagggcagcg gcactgcaga
     1381 ggtggagctg aagaagggag ccactctcaa aatcacgctg gataacgcct acatggaaaa
     1441 gtgtgacgag aacatcctgt ggctggacta caagaacatc tgcaaggtgg tggaagtggg
     1501 cagcaagatc tacgtggatg atgggcttat ttctctccag gtgaagcaga aaggtgccga
     1561 cttcctggtg acggaggtgg aaaatggtgg ctccttgggc agcaagaagg gtgtgaacct
     1621 tcctggggct gctgtggact tgcctgctgt gtcggagaag gacatccagg atctgaagtt
     1681 tggggtcgag caggatgttg atatggtgtt tgcgtcattc atccgcaagg catctgatgt
     1741 ccatgaagtt aggaaggtcc tgggagagaa gggaaagaac atcaagatta tcagcaaaat
     1801 cgagaatcat gagggggttc ggaggtttga tgaaatcctg gaggccagtg atgggatcat
     1861 ggtggctcgt ggtgatctag gcattgagat tcctgcagag aaggtcttcc ttgctcagaa
     1921 gatgatgatt ggacggtgca accgagctgg gaagcctgtc atctgtgcta ctcagatgct
     1981 ggagagcatg atcaagaagc cccgccccac tcgggctgaa ggcagtgatg tggccaatgc
     2041 agtcctggat ggagccgact gcatcatgct gtctggagaa acagccaaag gggactatcc
     2101 tctggaggct gtgcgcatgc agcacctgat tgcccgtgag gcagaggctg ccatctacca
     2161 cttgcaatta tttgaggaac tccgccgcct ggcgcccatt accagcgacc ccacagaagc
     2221 caccgccgtg ggtgccgtgg aggcctcctt caagtgctgc agtggggcca taatcgtcct
     2281 caccaagtct ggcaggtctg ctcaccaggt ggccagatac cgcccacgtg cccccatcat
     2341 tgctgtgacc cggaatcccc agacagctcg tcaggcccac ctgtaccgtg gcatcttccc
     2401 tgtgctgtgc aaggacccag tccaggaggc ctgggctgag gacgtggacc tccgggtgaa
     2461 ctttgccatg aatgttggca aggcccgagg cttcttcaag aagggagatg tggtcattgt
     2521 gctgaccgga tggcgccctg gctccggctt caccaacacc atgcgtgttg ttcctgtgcc
     2581 gaagcttggg cccgaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca
     2641 tcatcatcat catcattgag tttaaacggt ctccagctta agtttaaacc gctgatcagc
     2701 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2761 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2821 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2881 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2941 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3181 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3301 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3421 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3481 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3601 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3661 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3721 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3781 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3841 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3901 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3961 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4021 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4081 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4141 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4201 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4261 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4321 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4381 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4441 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4501 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4561 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4621 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4681 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4741 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4801 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4861 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4921 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4981 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5041 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5101 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5161 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5221 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5281 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5341 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5401 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5461 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5521 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5581 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5641 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5701 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5761 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5821 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5881 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7081 catttccccg aaaagtgcca cctgacgtc
//
