LOCUS       V019342                 6791 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019342
VERSION     V019342
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..1471
                     /label="RTP1(NM_153708)"
                     /note="RTP1(NM_153708)"
                     /gene="RTP1"
     CDS             1478..1543
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1567..2155
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2221..2269
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2471..2899
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2913..3242
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3309..4103
                     /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    4279..4400
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4449..4465)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4473..4489
                     /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(4497..4527)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4542..4563
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4851..5439)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5610..6470)
                     /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(6471..6575)
                     /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 ccatgaggat ttttagaccg tggagactgc gctgccctgc
      721 cctgcaccta ccctcactct ccgtgttctc actaaggtgg aaattgcctt ccctcactac
      781 tgacgagacc atgtgtaaaa gcgtgaccac agatgagtgg aagaaagtct tctatgagaa
      841 gatggaggag gcaaagccgg ctgacagctg ggacctcatc atagacccca acctcaagca
      901 caatgtgctg agccctggtt ggaagcagta cctggaattg catgcttcag gcaggttcca
      961 ctgctcctgg tgctggcaca cctggcagtc gccctacgtg gtcatcctct tccacatgtt
     1021 cctggaccgc gcccagcggg cgggctcggt gcgcatgcgc gtcttcaagc agctgtgcta
     1081 tgagtgcggc acggcgcggc tggacgagtc cagcatgctg gaggagaaca tcgagggcct
     1141 ggtggacaac ctcatcacca gcctgcgcga gcagtgctac ggcgagcgtg gcggccagta
     1201 ccgcatccac gtggccagcc gccaggacaa ccggcggcac cgcggagagt tctgcgaggc
     1261 ctgccaggag ggcatcgtgc actggaagcc cagcgagaag ctgctggagg aggaggcgac
     1321 cacctacacc ttctcccggg cgcccagccc caccaagtcg caggaccaga cgggctcagg
     1381 ctggaacttc tgctctatcc cctggtgctt gttttgggcc acggtcctgc tgctgatcat
     1441 ctacctgcag ttctctttcc gtagctccgt actcgaggac tacaaagacc atgacggtga
     1501 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat
     1561 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1621 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1681 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1741 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1801 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1861 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1921 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1981 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2041 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2101 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     2161 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2221 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     2281 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     2341 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     2401 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     2461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     2641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2821 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     2881 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3061 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3121 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3181 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3241 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4141 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4201 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4261 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4321 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4381 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4441 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4501 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4561 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4621 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4681 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4741 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5041 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6601 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     6661 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6721 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6781 atctgcttag g
//
