LOCUS       V031959                 7611 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031959
VERSION     V031959
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..2291
                     /label="Gsdme(NM_018769)"
                     /note="Gsdme(NM_018769)"
                     /gene="Gsdme"
     CDS             2298..2363
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2387..2975
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3041..3089
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3291..3719
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3733..4062
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4129..4923
                     /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    5099..5220
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5269..5285)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5293..5309
                     /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(5317..5347)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5362..5383
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5671..6259)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6430..7290)
                     /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(7291..7395)
                     /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 ccaccatgtt tgccaaagca actcgaaatt
      781 ttcttaaaga agttgatgct ggaggagacc tgatttcagt ctcacacttg aacgactctg
      841 acaagctgca acttctaagt ctggtgacca aaaagaagag atactggtgc tggcagagac
      901 ccaagtacca gattttatct gccaccctgg aagatgtact cacagaaggg cactgtctca
      961 gtccagtggt tgtggagtca gacttcgtga aatacgagag caagtgtgag aaccataaga
     1021 gcggggctat tgggacagtc gtggggaagg tcaagctgaa cgttggtggc aaaggcgtgg
     1081 tggagagtca ctcttcgttt ggaaccctga ggaagcagga ggtggacgtg cagcagctca
     1141 tccaggatgc cgtcaagaga acagttaata tggacaacct ggtacttcag caggtgctag
     1201 agagcaggaa cgaggtcctg tgtgtgctga cgcagaagat catgaccacg cagaagtgcg
     1261 tgatttctga gcatgtgcag tcggaggaga cgtgtggagg catggtgggg atccagacca
     1321 agactataca ggtgtcagca acggaggatg ggacggtcac cacggacacc aatgtagtgc
     1381 tggagatccc tgctgccacc accattgcct atggcatcat ggagctgttt gtgaaacaag
     1441 atggccagtt tgaattctgc ctcctccaag ggaaacatgg tggcttcgag catgagagga
     1501 aactagactc tgtctacttg gaccccctgg cctacagaga gttcgccttt ctggacatgc
     1561 tggatggggg tcaagggatc tcttcgcagg acgggcccct acgagttgta aaacaagcaa
     1621 ccctgcacct agagaggagt ttccatccct ttgcggtgtt acctgcccag cagcagaggg
     1681 cgctgttctg tgtcctgcag aaaatcctgt ttgatgaaga actccttcgg gccctggagc
     1741 aagtgtgtga tgatgtggct ggtggtctct ggtcctcaca ggctgtattg gcaatggagg
     1801 agctgaccga cagtcagcag caggacctca cagccttctt gcagctggtg ggatacagga
     1861 tacaaggaga gcatcctggc ccacaggatg aggtcagcaa ccagaagctc tttgcaacag
     1921 cctacttcct ggtcagcgca ctagcagaaa tgcctgataa tgccacagtt ttcctgggga
     1981 cttgctgcaa actccatgtt atttcttcgc tgtgctgctt gctccatgct ctgtctgatg
     2041 acagcgtgtg tgattttcac aaccccacct tggctcctct gagagacaca gagaggtttg
     2101 gcatcgtgca gcgattgttt gcctctgctg acattgccct ggagaggatg cagttttctg
     2161 cgaaagccac catcctgaag gactcttgca tcttcccgct aattcttcac atcactctaa
     2221 gtgggctcag cactctaagc aaagagcatg aggaagagct ttgtcagtca ggacatgcta
     2281 caggtcaaga cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2341 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2521 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2581 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2641 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2701 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2761 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2821 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2881 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2941 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3001 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3061 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3121 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3181 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3241 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3421 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3661 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3721 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3781 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3901 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3961 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4021 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4081 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4141 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4201 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4261 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4321 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4381 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4441 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4501 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4561 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4621 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4681 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4741 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4801 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4861 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4921 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4981 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5041 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5101 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5161 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5221 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5281 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5341 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5401 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5461 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5521 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5581 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5641 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5701 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5761 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5821 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5881 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5941 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6001 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6061 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6121 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6181 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6241 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6301 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6361 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6421 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6481 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6541 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6601 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6661 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6721 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6781 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6841 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6901 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6961 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7021 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7081 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7141 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7201 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7261 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7321 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7381 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7441 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7501 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7561 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
