LOCUS       V043201                 8237 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043201
VERSION     V043201
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..2143
                     /label="Gsdmd(NM_026960)"
                     /note="Gsdmd(NM_026960)"
                     /gene="Gsdmd"
     CDS             2180..2896
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             2903..2929
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2933..2959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2963..2989
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3013..3601
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3667..3715
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3917..4345
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4359..4688
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4755..5549
                     /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    5725..5846
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5895..5911)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5919..5935
                     /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(5943..5973)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5988..6009
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6297..6885)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7056..7916)
                     /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(7917..8021)
                     /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 ccatgccatc ggcctttgag aaagtggtca agaatgtgat
      721 caaggaggta agcggcagca gaggcgatct cattccggtg gacagcctgc ggaactccac
      781 cagcttcagg ccctactgcc ttctgaacag gaaattttca agctcaaggt tctggaaacc
      841 ccgttattca tgtgtcaacc tgtcaatcaa ggacatcctg gagcccagtg ctccagaacc
      901 agaaccggag tgttttggct ccttcaaagt ctctgatgtc gtcgatggga acattcaggg
      961 cagagtgatg ttgtcaggca tgggagaagg gaaaatttct ggtggggctg cagtgtctga
     1021 cagttccagt gcctccatga atgtgtgtat actgcgtgtg actcagaaga cctgggagac
     1081 catgcagcat gaaaggcacc ttcagcagcc tgagaacaaa atcctgcaac agcttcggag
     1141 tcgtggggat gacctgtttg tggtgaccga ggtgctgcag acaaaggagg aagtgcagat
     1201 cactgaggtc cacagccaag agggctcagg ccagtttacg ctgcctggag ctttatgctt
     1261 gaagggtgaa ggcaagggcc accaaagccg gaagaagatg gtgaccattc ctgcaggcag
     1321 catcctggca ttccgagtgg cccaactgct tattggctct aaatgggata tccttctcgt
     1381 ctcagatgag aaacagagga cctttgagcc ctcctcaggt gacagaaaag cagtgggcca
     1441 gaggcaccat ggcctcaatg tgcttgctgc gctttgttcc atcggaaagc agctcagtct
     1501 cctgtcagat gggattgatg aggaggaatt aattgaggcg gcagacttcc agggcctgta
     1561 tgctgaggtg aaggcttgct cctcagaact ggagagcttg gaaatggagt tgagacaaca
     1621 gatactggtg aacatcggaa agattttaca ggaccagccc agcatggaag ccttagaggc
     1681 ctcactaggg cagggcctgt gcagtggcgg ccaggtggag cctctggacg gcccagctgg
     1741 ctgcatcctt gagtgtctgg tgcttgactc tggagaactg gtgccggaac tcgcagcccc
     1801 tatcttctac ctgctgggag cactggctgt gctgagtgaa acccagcagc agctgctagc
     1861 taaggctctg gagacaacgg tgctgtcaaa gcagctggag ttggtgaagc acgtcttgga
     1921 acagagcacc ccgtggcagg agcagagttc tgtgtccctg cccaccgtgc tccttgggga
     1981 ctgctgggat gaaaagaatc ccacctgggt cttgctagaa gaatgtggcc taaggctgca
     2041 ggtagaatcc ccccaggtgc actgggaacc aacgtctctg atccccacaa gtgcgctcta
     2101 tgcctccctg ttcctattgt caagtctagg ccagaaacct tgtggatccg gcggtggagg
     2161 aagcggcgga ggtggatcta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2221 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2281 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2341 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2401 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2461 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2521 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2581 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2641 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2701 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2761 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2821 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2881 ggacgagctg tacaagctcg agtacccata cgacgtacct gattatgcaa cttatcctta
     2941 tgatgtacct gattatgcta gctatcctta tgatgtccca gactacgcat agtgagggcc
     3001 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3061 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3121 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3181 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3241 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3301 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3361 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3421 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3481 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3541 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3601 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3661 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     3721 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3781 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3841 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3901 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3961 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4021 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     4081 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4141 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4201 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4261 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     4321 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4381 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4441 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4561 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4621 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4681 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4741 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4801 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4861 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4921 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4981 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5041 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5101 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5161 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5221 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5281 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5341 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5401 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5461 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5521 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     5581 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5641 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5701 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5761 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5821 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5881 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5941 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6001 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6481 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7921 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8041 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     8101 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8161 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8221 tgaagaatct gcttagg
//
