LOCUS       V045978                 6531 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045978
VERSION     V045978
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..1211
                     /label="Ripply3(NM_133229)"
                     /note="Ripply3(NM_133229)"
                     /gene="Ripply3"
     CDS             1218..1283
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1307..1895
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1961..2009
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2211..2639
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2653..2982
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3049..3843
                     /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    4019..4140
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4189..4205)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4213..4229
                     /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(4237..4267)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4282..4303
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4591..5179)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5350..6210)
                     /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(6211..6315)
                     /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 ccaccatgag accggaggcg gccggagtcc
      781 gggaggcgcg gggacgcctc tgccactgtc cgggggatga tcccgggagg ctaccgctgc
      841 cacgcgggcc cgaaagttcc attccagccc cgtggagacc gtggatgtcg cctccccccg
      901 gagatgctga actgaccaga actgaacgtc cgtgtgagtc ttggggagac caacacacat
      961 cgggatcaaa gggagccttt ggattccagc atcctgtcag actttactta cccgtttcaa
     1021 agcgtcaaga ataccttcag agttctgggg aaaaagtgct ggccagcttc ccagtgcaag
     1081 ccaccattca tttctacaac gatgactctg agtctggctc tgaggaggaa caagaggagg
     1141 aggcccagcc caaccacctt cagtgcctgg aggcagaagt cagggattct gctcaggagg
     1201 aaagggcaga aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     1261 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     1321 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1381 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1441 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     1501 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     1561 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     1621 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1681 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     1741 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     1801 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     1861 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     1921 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     1981 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2041 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     2101 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     2161 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     2221 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2281 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2341 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     2401 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2461 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2521 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2581 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2641 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2701 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2821 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2881 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2941 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3001 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3061 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3121 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3181 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3241 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3301 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3361 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3421 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3481 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3541 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3601 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3661 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3721 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3781 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3841 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3901 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3961 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4141 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4201 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4261 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4321 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6301 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6361 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     6421 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6481 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
