LOCUS       V021197                 7523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021197
VERSION     V021197
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..2170
                     /label="GSDME(NM_001127453)"
                     /note="GSDME(NM_001127453)"
                     /gene="GSDME"
     CDS             2210..2275
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2299..2887
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2953..3001
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3203..3631
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3645..3974
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4041..4835
                     /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    5011..5132
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5181..5197)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5205..5221
                     /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(5229..5259)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5274..5295
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5583..6171)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6342..7202)
                     /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(7203..7307)
                     /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 ccatgtttgc caaagcaacc aggaattttc ttagagaagt
      721 tgatgctgat ggtgacctga ttgcagtatc aaatctgaat gactctgata agttacagct
      781 tctaagtctg gtgacaaaaa agaagagatt ctggtgctgg cagagaccca agtaccagtt
      841 tttatccctc acccttggcg atgtactcat agaagaccaa tttccgagtc cagtggtcgt
      901 ggagtcggac tttgtgaaat acgagggcaa gtttgcaaac cacgtgagtg gaaccctgga
      961 gactgcactg gggaaggtca agctgaacct ggggggcagc agccgcgtag agagccagtc
     1021 ttcatttgga accctgagga agcaggaggt ggatttgcag cagctcatca gagactctgc
     1081 cgagagaaca ataaatctga gaaaccctgt gctccagcag gtgctggaag gaaggaatga
     1141 ggtcctgtgc gttttgacac agaagatcac gacgatgcag aagtgtgtga tctctgagca
     1201 catgcaggtc gaggagaagt gtggtggcat cgtgggcatc cagaccaaga cggtgcaggt
     1261 gtcagcgacg gaggatggga atgtcaccaa ggactccaac gtggtgctgg agatcccagc
     1321 tgccaccacc attgcctacg gtgtcattga gttatacgtg aaactggacg gccagttcga
     1381 gttctgcctt ctccgaggga agcaaggtgg cttcgagaac aagaagagaa ttgactctgt
     1441 ctacctggac cccctggtct ttcgagagtt tgcattcata gacatgccag atgctgcgca
     1501 tgggatatct tcccaggatg gaccattaag tgttttaaag caagcgaccc tgctcctgga
     1561 gaggaatttc catccatttg cggagctgcc tgagccacaa cagacagctt tgagtgacat
     1621 cttccaggcg gtcctatttg atgatgaact actcatggtc ctggaaccag tgtgcgatga
     1681 cctggtcagc ggcctctcgc ccacagtggc ggtgctgggg gagctgaagc cccggcagca
     1741 gcaggacctt gtggccttcc tgcagctggt ggggtgcagc ttacagggtg ggtgtccggg
     1801 ccccgaggat gcaggcagca agcagctgtt tatgacagcc tacttcttgg tcagtgccct
     1861 cgcagaaatg ccagatagcg cagcagctct gctgggcact tgctgcaaac tccagatcat
     1921 tcccacactg tgccacttgc ttcgtgctct gtctgatgat ggagtatctg atcttgaaga
     1981 cccaaccttg actcccctga aagatacaga aaggtttggg attgtgcagc gcttgtttgc
     2041 ctcagctgac attagtctgg agagactgaa gtcatctgtg aaagctgtca ttctgaagga
     2101 ctctaaagtc ttcccactgc ttctttgtat aaccctgaat ggactctgtg ctttaggcag
     2161 agaacattca gaattctgca gatatccagc acagtggcgg ccgctcgagg actacaaaga
     2221 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     2281 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2341 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2401 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2461 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2521 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2581 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2641 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     2701 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2761 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2821 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2881 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2941 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3001 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3061 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3121 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     3361 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3541 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3901 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4861 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4921 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4981 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5041 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5101 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5161 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5221 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5281 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5341 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5401 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5461 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5521 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5761 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6001 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7321 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     7381 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7441 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7501 attgcatgaa gaatctgctt agg
//
