LOCUS       V030654                10523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030654
VERSION     V030654
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"
     CDS             683..5182
                     /label="CPS1(NM_001875)"
                     /note="CPS1(NM_001875)"
                     /gene="CPS1"
     CDS             5189..5215
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5219..5245
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             5249..5275
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    5299..5887
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5953..6001
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6203..6631
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6645..6974
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7041..7835
                     /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    8011..8132
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8181..8197)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8205..8221
                     /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(8229..8259)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8274..8295
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8583..9171)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9342..10202)
                     /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(10203..10307)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgacgag gattttgaca gctttcaaag tggtgaggac
      721 actgaagact ggttttggct ttaccaatgt gactgcacac caaaaatgga aattttcaag
      781 acctggcatc aggctccttt ctgtcaaggc acagacagca cacattgtcc tggaagatgg
      841 aactaagatg aaaggttact cctttggcca tccatcctct gttgctggtg aagtggtttt
      901 taatactggc ctgggagggt acccagaagc tattactgac cctgcctaca aaggacagat
      961 tctcacaatg gccaacccta ttattgggaa tggtggagct cctgatacta ctgctctgga
     1021 tgaactggga cttagcaaat atttggagtc taatggaatc aaggtttcag gtttgctggt
     1081 gctggattat agtaaagact acaaccactg gctggctacc aagagtttag ggcaatggct
     1141 acaggaagaa aaggttcctg caatttatgg agtggacaca agaatgctga ctaaaataat
     1201 tcgggataag ggtaccatgc ttgggaagat tgaatttgaa ggtcagcctg tggattttgt
     1261 ggatccaaat aaacagaatt tgattgctga ggtttcaacc aaggatgtca aagtgtacgg
     1321 caaaggaaac cccacaaaag tggtagctgt agactgtggg attaaaaaca atgtaatccg
     1381 cctgctagta aagcgaggag ctgaagtgca cttagttccc tggaaccatg atttcaccaa
     1441 gatggagtat gatgggattt tgatcgcggg aggaccgggg aacccagctc ttgcagaacc
     1501 actaattcag aatgtcagaa agattttgga gagtgatcgc aaggagccat tgtttggaat
     1561 cagtacagga aacttaataa caggattggc tgctggtgcc aaaacctaca agatgtccat
     1621 ggccaacaga gggcagaatc agcctgtttt gaatatcaca aacaaacagg ctttcattac
     1681 tgctcagaat catggctatg ccttggacaa caccctccct gctggctgga aaccactttt
     1741 tgtgaatgtc aacgatcaaa caaatgaggg gattatgcat gagagcaaac ccttcttcgc
     1801 tgtgcagttc cacccagagg tcaccccggg gccaatagac actgagtacc tgtttgattc
     1861 ctttttctca ctgataaaga aaggaaaagc taccaccatt acatcagtct taccgaagcc
     1921 agcactagtt gcatctcggg ttgaggtttc caaagtcctt attctaggat caggaggtct
     1981 gtccattggt caggctggag aatttgatta ctcaggatct caagctgtaa aagccatgaa
     2041 ggaagaaaat gtcaaaactg ttctgatgaa cccaaacatt gcatcagtcc agaccaatga
     2101 ggtgggctta aagcaagcgg atactgtcta ctttcttccc atcacccctc agtttgtcac
     2161 agaggtcatc aaggcagaac agccagatgg gttaattctg ggcatgggtg gccagacagc
     2221 tctgaactgt ggagtggaac tattcaagag aggtgtgctc aaggaatatg gtgtgaaagt
     2281 cctgggaact tcagttgagt ccattatggc tacggaagac aggcagctgt tttcagataa
     2341 actaaatgag atcaatgaaa agattgctcc aagttttgca gtggaatcga ttgaggatgc
     2401 actgaaggca gcagacacca ttggctaccc agtgatgatc cgttccgcct atgcactggg
     2461 tgggttaggc tcaggcatct gtcccaacag agagactttg atggacctca gcacaaaggc
     2521 ctttgctatg accaaccaaa ttctggtgga gaagtcagtg acaggttgga aagaaataga
     2581 atatgaagtg gttcgagatg ctgatgacaa ttgtgtcact gtctgtaaca tggaaaatgt
     2641 tgatgccatg ggtgttcaca caggtgactc agttgttgtg gctcctgccc agacactctc
     2701 caatgccgag tttcagatgt tgagacgtac ttcaatcaat gttgttcgcc acttgggcat
     2761 tgtgggtgaa tgcaacattc agtttgccct tcatcctacc tcaatggaat actgcatcat
     2821 tgaagtgaat gccagactgt cccgaagctc tgctctggcc tcaaaagcca ctggctaccc
     2881 attggcattc attgctgcaa agattgccct aggaatccca cttccagaaa ttaagaacgt
     2941 cgtatccggg aagacatcag cctgttttga acctagcctg gattacatgg tcaccaagat
     3001 tccccgctgg gatcttgacc gttttcatgg aacatctagc cgaattggta gctctatgaa
     3061 aagtgtagga gaggtcatgg ctattggtcg tacctttgag gagagtttcc agaaagcttt
     3121 acggatgtgc cacccatcta tagaaggttt cactccccgt ctcccaatga acaaagaatg
     3181 gccatctaat ttagatctta gaaaagagtt gtctgaacca agcagcacgc gtatctatgc
     3241 cattgccaag gccattgatg acaacatgtc ccttgatgag attgagaagc tcacatacat
     3301 tgacaagtgg tttttgtata agatgcgtga tattttaaac atggaaaaga cactgaaagg
     3361 cctcaacagt gagtccatga cagaagaaac cctgaaaagg gcaaaggaga ttgggttctc
     3421 agataagcag atttcaaaat gccttgggct cactgaggcc cagacaaggg agctgaggtt
     3481 aaagaaaaac atccaccctt gggttaaaca gattgataca ctggctgcag aatacccatc
     3541 agtaacaaac tatctctatg ttacctacaa tggtcaggag catgatgtca attttgatga
     3601 ccatggaatg atggtgctag gctgtggtcc atatcacatt ggcagcagtg tggaatttga
     3661 ttggtgtgct gtctctagta tccgcacact gcgtcaactt ggcaagaaga cggtggtggt
     3721 gaattgcaat cctgagactg tgagcacaga ctttgatgag tgtgacaaac tgtactttga
     3781 agagttgtcc ttggagagaa tcctagacat ctaccatcag gaggcatgtg gtggctgcat
     3841 catatcagtt ggaggccaga ttccaaacaa cctggcagtt cctctataca agaatggtgt
     3901 caagatcatg ggcacaagcc ccctgcagat cgacagggct gaggatcgct ccatcttctc
     3961 agctgtcttg gatgagctga aggtggctca ggcaccttgg aaagctgtta atactttgaa
     4021 tgaagcactg gaatttgcaa agtctgtgga ctacccctgc ttgttgaggc cttcctatgt
     4081 tttgagtggg tctgctatga atgtggtatt ctctgaggat gagatgaaaa aattcctaga
     4141 agaggcgact agagtttctc aggagcaccc agtggtgctg acaaaatttg ttgaaggggc
     4201 ccgagaagta gaaatggacg ctgttggcaa agatggaagg gttatctctc atgccatctc
     4261 tgaacatgtt gaagatgcag gtgtccactc gggagatgcc actctgatgc tgcccacaca
     4321 aaccatcagc caaggggcca ttgaaaaggt gaaggatgct acccggaaga ttgcaaaggc
     4381 ttttgccatc tctggtccat tcaacgtcca atttcttgtc aaaggaaatg atgtcttggt
     4441 gattgagtgt aacttgagag cttctcgatc cttccccttt gtttccaaga ctcttggggt
     4501 tgacttcatt gatgtggcca ccaaggtgat gattggagag aatgttgatg agaaacatct
     4561 tccaacattg gaccatccca taattcctgc tgactatgtt gcaattaagg ctcccatgtt
     4621 ttcctggccc cggttgaggg atgctgaccc cattctgaga tgtgagatgg cttccactgg
     4681 agaggtggct tgctttggtg aaggtattca tacagccttc ctaaaggcaa tgctttccac
     4741 aggatttaag ataccccaga aaggcatcct gataggcatc cagcaatcat tccggccaag
     4801 attccttggt gtggctgaac aattacacaa tgaaggtttc aagctgtttg ccacggaagc
     4861 cacatcagac tggctcaacg ccaacaatgt ccctgccacc ccagtggcat ggccgtctca
     4921 agaaggacag aatcccagcc tctcttccat cagaaaattg attagagatg gcagcattga
     4981 cctagtgatt aaccttccca acaacaacac taaatttgtc catgataatt atgtgattcg
     5041 gaggacagct gttgatagtg gaatccctct cctcactaat tttcaggtga ccaaactttt
     5101 tgctgaagct gtgcagaaat ctcgcaaggt ggactccaag agtcttttcc actacaggca
     5161 gtacagtgct ggaaaagcag cactcgagta cccatacgac gtacctgatt atgcaactta
     5221 tccttatgat gtacctgatt atgctagcta tccttatgat gtcccagact acgcatagtg
     5281 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5341 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5401 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5461 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5521 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5581 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5641 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     5701 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5761 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5821 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5881 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5941 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     6001 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6061 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6121 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     6181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     6361 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6541 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     6601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     6661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     6841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6901 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     7021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     7081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     7141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     7201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     7261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     7321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     7381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     7441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     7501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     7561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     7621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     7681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     7741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     7801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     7861 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     7921 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     7981 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     8041 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     8101 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     8161 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     8221 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     8281 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     8341 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     8401 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     8461 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     8521 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8761 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     9001 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     9061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     9121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     9181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     9241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     9301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     9361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     9421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     9481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     9541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     9601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     9661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     9961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    10021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    10081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    10141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    10201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    10261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    10321 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
    10381 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
    10441 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
    10501 attgcatgaa gaatctgctt agg
//
