LOCUS       V030232                 8734 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030232
VERSION     V030232
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             706..3393
                     /label="Dsc3(NM_007882)"
                     /note="Dsc3(NM_007882)"
                     /gene="Dsc3"
     CDS             3400..3426
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3430..3456
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3460..3486
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3510..4098
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4164..4212
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4414..4842
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4856..5185
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5252..6046
                     /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    6222..6343
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6392..6408)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6416..6432
                     /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(6440..6470)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6485..6506
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6794..7382)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7553..8413)
                     /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(8414..8518)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggt ggttcctgag
      721 ttccggagcc cacagtgcag ggcactatgc acgaagctgc tgctgaccct ctgggtcttc
      781 agttttgttg gtgaagcctg taaaaaggtg acatttcatg taccttcaac actagaggca
      841 gacaagatca tcggaagagt tagtttgaaa gagtgtctca gctcggcaga tggcatcatg
      901 ccaagtgatc ctgatttcag agttctagat gatgggtcag tttatccgac cagagctgtt
      961 gtattgtctg atgagaaaag atcttttacc attcagcttt ctgattcaaa gatgcagaca
     1021 cagaaagaga tccctgtaat cctggagcac aagaagaagg tactgaagaa aagacacact
     1081 aaggaaactg tcctcagaag atctaagaga agatgggcac ccattccatg ttcaatgcaa
     1141 gaaaattcct tgggtccctt cccattgttt cttcaacagg ttcagtctga tgcagctcag
     1201 aactacacag tcttctattc aataagtggg cgtggagctg atcaagaacc acttaactgg
     1261 tttttcatcg agagagacac tgggaatcta tattgtactc ggcctgtgga tcgtgaggaa
     1321 tatgatgttt ttgatttgat tgcctatgca tccactgcag atggctattc agcagattta
     1381 cctctcccac tgcccatcaa aatagaggat gaaaatgaca attaccctct ttttacagaa
     1441 gcaatttatg cttttgaagt tccagaaggc agtagacttg gtactgtagt gggaacagtt
     1501 tgtgccacgg ataaagatga gcctgacact atgcacacac gcctcaagta cagcatcttg
     1561 gagcagaccc caccatcacc tgggcttttt tctgtgcacc ctgacacagg tgtcatcacc
     1621 accgtctctc actacatgga cagagaggtc gtggacaagt acaaactgat aatgaaagtt
     1681 caggacatga atggccaatt ttttggacta atcagtacat caacttgtat cataactgtg
     1741 caagattcca atgacaatgc acccacattc agacaaaata cttatgaaac tgctgtggag
     1801 gaaaatacat ataatgttga aatcttgcgc atacctgtag atgataagga catgattaac
     1861 actgccaact ggaaggcaaa tttcaccatc ttaaaaggaa atgaaaatgg atggttcaaa
     1921 atcacaacag acccagtgac aaatgaaggt gttctgtgtg ttgtgaagcc actggattat
     1981 gaagaaaatc gtcaagtgac cctagagatt ggtgtgaaca acgaagctcc attcattaaa
     2041 gatgttgcaa accgaattcc caccatgaac cgggctatgg ttacagtaca tgtgaaggat
     2101 cagaatgagg ggccagaatg caagccccca gagcaatatg tgcggatcaa agagaactca
     2161 gcagttgggt ctaagatcaa tggctacaag gcctacgacc ctgaaaccaa gaatagcaat
     2221 ggcttaaggt acaaaaaatt gcaggatcct aaagactggg tcagcattga agaagtttcg
     2281 gggttgctta ccatatcaaa aaccctagac agagaaataa tggctccaag aaatgacatg
     2341 tacaatatta cagtcatggc aattgaccaa gagggtaaat catgcacagg aacacttgca
     2401 gtgaacatcg aggatgtgaa tgacaacgca ccagagataa ttcaagatta tattgttatc
     2461 tgtaagccaa agatggggta cacggacatt tcagctgtgg atcctgatga acctatccat
     2521 ggccctccct ttcagttcaa cttggcaaac acttctccag aagtcaacag aatctggacc
     2581 ctcaaccaag ttaatgatac agctgctcgc ctgtcctatc agaaaactgc ggatgttcaa
     2641 atatacaacg ttcctgttac tgtaaaagac agagcaggcc aatctgcaac caaaatcttg
     2701 agagttaacc tgtgtgattg tactcatccc agccaatgtc cgctcagatc aaggagtgca
     2761 ggaattaccc tgggaaagtg ggccatcctt gccatcttgc tgggcattgc tctgctattt
     2821 tcggtactgc taactttggt atgtggagtt gttactgccc gaaaaggaaa acacttccct
     2881 gaagatttag cgcagcaaaa cttaattata tcaaataccg aagcacctgg ggatgacaga
     2941 gtgtgctctg ccaatggatt caccacccac actgccaaca actccagcca aggcttctgt
     3001 ggtacaatgg ggtcaggaat gaggaatggt gggcaggaga ccatcgagat gatgaaagga
     3061 caccagacct tggactcctg ccgtgtggcc gggcatcacc ataccctgga ctccggcagg
     3121 ggaggacaca tggatactga caactgcaga tacacatact cagagtggca cagtttcact
     3181 caaccccgac ttggtgaaaa attgcatgtg tgtaatcaga atgaagacca catcccatcc
     3241 caagactatg tcctcaccta taactatgaa ggcagagggt ctccggccgg ctctgtaggt
     3301 tgctgcagtg aaaagcagga agaagaggga cttgactttt taaacaactt ggaacccaaa
     3361 tttcttacat tggcagaaac atgcacaaag agactcgagt acccatacga cgtacctgat
     3421 tatgcaactt atccttatga tgtacctgat tatgctagct atccttatga tgtcccagac
     3481 tacgcatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     3541 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3601 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3661 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3721 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3781 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3841 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3901 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3961 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4021 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4081 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     4141 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     4201 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4261 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4321 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     4381 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4441 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4501 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4561 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4621 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4681 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4741 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4801 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4861 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4921 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4981 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5041 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5101 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5161 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5221 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5281 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5341 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5401 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5461 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5521 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5581 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5641 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5701 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5761 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5821 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5881 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5941 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6001 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6061 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6121 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6181 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6361 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7681 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7861 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8161 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8521 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8581 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     8641 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     8701 cttgaccgac aattgcatga agaatctgct tagg
//
