LOCUS       V042867                 7084 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042867
VERSION     V042867
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2523
                     /label="Pkm(NM_001253883)"
                     /note="Pkm(NM_001253883)"
                     /gene="Pkm"
     CDS             2530..2556
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2560..2586
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2590..2616
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2651..2875
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2921..3349
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3363..3692
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3759..4553
                     /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    4727..4848
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4897..4913)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4921..4937
                     /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(4945..4975)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4990..5011
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5299..5884)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6055..6915)
                     /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(6916..7020)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgccgaagc cacacagtga agcagggact
      961 gccttcattc agacccagca gctccatgca gccatggctg acaccttcct ggaacacatg
     1021 tgccgcctgg acattgactc tgcccccatc acggcccgca acactggcat catttgtacc
     1081 attgggcctg cttcccgatc tgtggagatg ctgaaggaga tgattaagtc tggaatgaat
     1141 gtggctcggc tgaatttctc tcatggaacc catgagtacc atgcagagac catcaagaat
     1201 gtccgtgaag ccacagaaag ctttgcatct gatcccattc tctaccgtcc tgttgcggtg
     1261 gctctggata caaagggacc tgagatccgg actggactca tcaagggcag cggcaccgct
     1321 gaggtggagc tgaagaaggg agccactctg aagatcaccc tggacaacgc ttacatggag
     1381 aagtgtgacg agaacatcct gtggctggac tacaagaaca tctgcaaggt ggtggaggtg
     1441 ggcagcaaga tctacgtgga cgatgggctc atctcactgc aggtgaagga gaaaggcgct
     1501 gacttcctgg tgacggaggt ggagaatggt ggctccttgg gcagcaagaa gggcgtgaac
     1561 ctgccgggcg ctgctgtgga tctccccgct gtgtcggaaa aggacatcca ggacctgaag
     1621 tttggggtgg agcaggatgt ggacatggtg tttgcatctt tcatccgcaa ggcagccgac
     1681 gtgcatgaag tcaggaaggt gctgggagag aagggcaaga acatcaagat catcagcaaa
     1741 atcgagaacc atgaaggcgt ccgcaggttt gatgagatct tggaggccag tgatgggatc
     1801 atggtggctc gtggtgacct gggcattgag attcctgcag agaaggtctt cctggctcag
     1861 aagatgatga tcgggcgatg caaccgagct gggaagcctg tcatctgtgc cacacagatg
     1921 ctggagagca tgatcaagaa gccacgcccc acccgtgctg aaggcagtga tgtggccaat
     1981 gcagtcctgg atggagcaga ctgcatcatg ctgtctggag aaacagccaa gggggactac
     2041 cctctggagg ctgttcgcat gcagcacctg atagctcggg aggctgaggc agccatgttc
     2101 caccgtctgc tgtttgaaga gcttgtgcga gcctccagtc actccacaga cctcatggag
     2161 gccatggcca tgggcagcgt ggaggcctct tataagtgtt tagcagcagc tttgatagtt
     2221 ctcacggagt ctggcaggag tgctcaccaa gtggccaggt accgccctcg ggctcctatc
     2281 attgccgtga ctcgaaatcc ccagactgct cgccaggccc atctgtaccg tggcatcttc
     2341 cctgtgctgt gtaaggatgc cgtgctgaat gcctgggctg aggatgtcga ccttcgtgta
     2401 aacttggcca tggatgttgg caaggcccga ggcttcttca agaagggaga tgtggtcatt
     2461 gtgctgaccg ggtggcgccc tggctctgga ttcaccaaca ccatgcgtgt agtgcctgta
     2521 cctctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     2581 tatgctagct atccttatga tgtcccagac tacgcatgag ggcccgttta aacccgctga
     2641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2761 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2821 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2881 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3301 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3361 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3421 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3541 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3601 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3661 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3721 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3781 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3841 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3901 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3961 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4021 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4081 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4141 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4201 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4261 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4321 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4381 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4441 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4501 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4561 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4621 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4681 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4741 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4801 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4861 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4921 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4981 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5041 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5101 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5161 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5221 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5281 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5341 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5401 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5461 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5521 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5581 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5641 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5701 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5761 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5821 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5881 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5941 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6001 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6061 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6121 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6181 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6241 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6301 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6361 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6421 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6481 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6541 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6601 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6661 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6721 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6781 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6841 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6901 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6961 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7021 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7081 tggt
//
