LOCUS       V044237                 7641 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044237
VERSION     V044237
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..2321
                     /label="Pnkp(NM_021549)"
                     /note="Pnkp(NM_021549)"
                     /gene="Pnkp"
     CDS             2328..2393
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2417..3005
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3071..3119
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3321..3749
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3763..4092
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4159..4953
                     /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    5129..5250
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5299..5315)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5323..5339
                     /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(5347..5377)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5392..5413
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5701..6289)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6460..7320)
                     /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(7321..7425)
                     /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 ccaccatgtc acagcttgga tcccggggcc
      781 gcctgtggct gcagagcccc accggtgggc cgccccctat attcctgccg tcggatggcc
      841 aagccctggt cctcggtcgg ggacccctga cccaagtgac ggaccggaag tgctcccgaa
      901 accaagtgga gctgattgca gaccctgaaa gccggacagt ggcagtgaaa cagctgggtg
      961 ttaacccgtc aactgttggg gtccaggagt tgaagccagg actgtctggc tctctaagct
     1021 tgggggatgt cctttatctg gttaatggcc tatatcccct gaccctgcgt tgggaagagc
     1081 tcagcacatc aggatcccag ccagatgctc caccagacac tcctggggat ccggaggagg
     1141 gagaggatac tgagcctcag aaaaagcgag ttcggaaatc ctctcttggc tgggagagct
     1201 taaagaagtt actggtgttc acagcgtctg gggtgaagcc ccagggcaag gtggctgcct
     1261 ttgacctaga tgggaccctc atcaccaccc gctctggaaa ggtcttcccc actagcccta
     1321 gtgactggag gattctgtac ccagagattc caaagaagct ccaggagctg gctgctgaag
     1381 gctacaagtt ggtaatcttc accaaccaaa tgggcatcgg gcgaggcaag ctgccagcag
     1441 aggtgttcaa gggcaaggtg gaagctgtat tggagaagtt aggggtccca tttcaggtgc
     1501 tggtggcaac acatgcaggt ctaaaccgaa agccagtgag tggcatgtgg gaccatctgc
     1561 aggagcaggc caacgagggc atccccatct cggtagagga cagtgtcttc gtgggagatg
     1621 cagcagggcg cctagccaac tgggcaccgg ggaggaaaaa aaaggatttc tcttgtgcag
     1681 accgcctgtt cgcccttaac gtcggcctgc cctttgccac tccggaagag tttttcctta
     1741 agtggccggc tgcccgcttt gagcttccag ctttcgaccc gaggactatc tccagtgctg
     1801 ggcccctcta ccttccagag tccagctctc tcctgagccc caacccagag gtggtggtcg
     1861 cagtaggatt ccctggggcc ggcaagtcca ctttcatcca ggagcaccta gtgtcagctg
     1921 gctacgtcca tgtgaacagg gacacgctgg gctcgtggca gcgctgtgtg agctcatgcc
     1981 aggcggcact gaggcagggg aagcgagtcg tgattgacaa caccaaccca gatgtcccaa
     2041 gtcgggccag gtacatccaa tgtgccaaag atgcaggtgt gccctgccgc tgtttcaact
     2101 tctgtgctac aatagagcag gcacgccaca acaacaggtt tcgtgagatg accgaccctt
     2161 cgcatgcacc tgtgtcggac atggtcatgt ttagctacag gaagcagttt gagccgccca
     2221 cactggctga aggcttcctg gagatccttg agattccatt ccggttacag gagcatctgg
     2281 acccggcact gcagcgactg taccgccagt tttccgaggg cggatccgac tacaaagacc
     2341 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     2401 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2461 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2521 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2581 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2641 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2701 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2761 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     2821 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2881 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2941 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3001 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3061 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     3121 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3181 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3241 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     3301 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3361 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3421 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3481 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3541 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3601 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3661 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3721 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3781 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3841 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3901 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3961 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4021 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4081 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4141 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4201 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4261 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4321 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4381 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4441 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4501 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4561 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4621 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4681 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4741 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4801 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4861 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4921 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4981 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5041 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5101 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5161 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5221 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5281 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5341 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5401 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5461 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5881 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6121 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7441 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7501 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7561 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7621 tgcatgaaga atctgcttag g
//
