LOCUS       V022098                 8957 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022098
VERSION     V022098
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..3583
                     /label="ANPEP(NM_001150)"
                     /note="ANPEP(NM_001150)"
                     /gene="ANPEP"
     CDS             3623..3649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3653..3679
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3683..3709
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3733..4321
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4387..4435
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4637..5065
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5079..5408
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5475..6269
                     /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    6445..6566
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6615..6631)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6639..6655
                     /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(6663..6693)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6708..6729
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7017..7605)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7776..8636)
                     /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(8637..8741)
                     /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 ccatggccaa gggcttctat atttccaagt ccctgggcat
      721 cctggggatc ctcctgggcg tggcagccgt gtgcacaatc atcgcactgt cagtggtgta
      781 ctcccaggag aagaacaaga acgccaacag ctcccccgtg gcctccacca ccccgtccgc
      841 ctcagccacc accaaccccg cctcggccac caccttggac caaagtaaag cgtggaatcg
      901 ttaccgcctc cccaacacgc tgaaacccga ttcctaccgg gtgacgctga gaccgtacct
      961 cacccccaat gacaggggcc tgtacgtttt taagggctcc agcaccgtcc gtttcacctg
     1021 caaggaggcc actgacgtca tcatcatcca cagcaagaag ctcaactaca ccctcagcca
     1081 ggggcacagg gtggtcctgc gtggtgtggg aggctcccag ccccccgaca ttgacaagac
     1141 tgagctggtg gagcccaccg agtacctggt ggtgcacctc aagggctccc tggtgaagga
     1201 cagccagtat gagatggaca gcgagttcga gggggagttg gcagatgacc tggcgggctt
     1261 ctaccgcagc gagtacatgg agggcaatgt cagaaaggtg gtggccacta cacagatgca
     1321 ggctgcagat gcccggaagt ccttcccatg cttcgatgag ccggccatga aggccgagtt
     1381 caacatcacg cttatccacc ccaaggacct gacagccctg tccaacatgc ttcccaaagg
     1441 tcccagcacc ccacttccag aagaccccaa ctggaatgtc actgagttcc acaccacgcc
     1501 caagatgtcc acgtacttgc tggccttcat tgtcagtgag ttcgactacg tggagaagca
     1561 ggcatccaat ggtgtcttga tccggatctg ggcccggccc agtgccattg cggcgggcca
     1621 cggcgattat gccctgaacg tgacgggccc catccttaac ttctttgctg gtcattatga
     1681 cacaccctac ccactcccaa aatcagacca gattggcctg ccagacttca acgccggcgc
     1741 catggagaac tggggactgg tgacctaccg ggagaactcc ctgctgttcg accccctgtc
     1801 ctcctccagc agcaacaagg agcgggtggt cactgtgatt gctcatgagc tggcccacca
     1861 gtggttcggg aacctggtga ccatagagtg gtggaatgac ctgtggctga acgagggctt
     1921 cgcctcctac gtggagtacc tgggtgctga ctatgcggag cccacctgga acttgaaaga
     1981 cctcatggtg ctgaatgatg tgtaccgcgt gatggcagtg gatgcactgg cctcctccca
     2041 cccgctgtcc acacccgcct cggagatcaa cacgccggcc cagatcagtg agctgtttga
     2101 cgccatctcc tacagcaagg gcgcctcagt cctcaggatg ctctccagct tcctgtccga
     2161 ggacgtattc aagcagggcc tggcgtccta cctccacacc tttgcctacc agaacaccat
     2221 ctacctgaac ctgtgggacc acctgcagga ggctgtgaac aaccggtcca tccaactccc
     2281 caccaccgtg cgggacatca tgaaccgctg gaccctgcag atgggcttcc cggtcatcac
     2341 ggtggatacc agcacgggga ccctttccca ggagcacttc ctccttgacc ccgattccaa
     2401 tgttacccgc ccctcagaat tcaactacgt gtggattgtg cccatcacat ccatcagaga
     2461 tggcagacag cagcaggact actggctgat agatgtaaga gcccagaacg atctcttcag
     2521 cacatcaggc aatgagtggg tcctgctgaa cctcaatgtg acgggctatt accgggtgaa
     2581 ctacgacgaa gagaactgga ggaagattca gactcagctg cagagagacc actcggccat
     2641 ccctgtcatc aatcgggcac agatcattaa tgacgccttc aacctggcca gtgcccataa
     2701 ggtccctgtc actctggcgc tgaacaacac cctcttcctg attgaagaga gacagtacat
     2761 gccctgggag gccgccctga gcagcctgag ctacttcaag ctcatgtttg accgctccga
     2821 ggtctatggc cccatgaaga actacctgaa gaagcaggtc acacccctct tcattcactt
     2881 cagaaataat accaacaact ggagggagat cccagaaaac ctgatggacc agtacagcga
     2941 ggttaatgcc atcagcaccg cctgctccaa cggagttcca gagtgtgagg agatggtctc
     3001 tggccttttc aagcagtgga tggagaaccc caataataac ccgatccacc ccaacctgcg
     3061 gtccaccgtc tactgcaacg ctatcgccca gggcggggag gaggagtggg acttcgcctg
     3121 ggagcagttc cgaaatgcca cactggtcaa tgaggctgac aagctccggg cagccctggc
     3181 ctgcagcaaa gagttgtgga tcctgaacag gtacctgagc tacaccctga acccggactt
     3241 aatccggaag caggacgcca cctctaccat catcagcatt accaacaacg tcattgggca
     3301 aggtctggtc tgggactttg tccagagcaa ctggaagaag ctttttaacg attatggtgg
     3361 tggctcgttc tccttctcca acctcatcca ggcagtgaca cgacgattct ccaccgagta
     3421 tgagctgcag cagctggagc agttcaagaa ggacaacgag gaaacaggct tcggctcagg
     3481 cacccgggcc ctggagcaag ccctggagaa gacgaaagcc aacatcaagt gggtgaagga
     3541 gaacaaggag gtggtgctcc agtggttcac agaaaacagc aaagaattct gcagatatcc
     3601 agcacagtgg cggccgctcg agtacccata cgacgtacct gattatgcaa cttatcctta
     3661 tgatgtacct gattatgcta gctatcctta tgatgtccca gactacgcat agtgagggcc
     3721 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3781 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3841 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3901 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3961 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4021 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4081 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4141 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4201 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4261 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4321 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4381 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     4441 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4501 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4561 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     4621 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4681 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4741 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     4801 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4861 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4921 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4981 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     5041 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5101 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5161 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5221 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5281 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5341 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5401 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5461 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5521 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5581 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5641 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5701 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5761 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5821 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5881 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5941 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6001 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6061 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6121 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6181 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6241 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6301 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6361 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6421 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6481 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6541 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6601 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6661 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7201 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7441 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8761 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     8821 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8881 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8941 tgaagaatct gcttagg
//
