LOCUS       V018291                 6839 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018291
VERSION     V018291
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..2325
                     /label="GSDMD(NM_024736)"
                     /note="GSDMD(NM_024736)"
                     /gene="GSDMD"
     CDS             2326..2352
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2406..2630
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2676..3104
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3118..3447
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3514..4308
                     /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    4482..4603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4652..4668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4676..4692
                     /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(4700..4730)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4745..4766
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5054..5639)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5810..6670)
                     /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(6671..6775)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggggt cggcctttga gcgggtagtc
      901 cggagagtgg tccaggagct ggaccatggt ggggagttca tccctgtgac cagcctgcag
      961 agctccactg gcttccagcc ctactgcctg gtggttagga agccctcaag ctcatggttc
     1021 tggaaacccc gttataagtg tgtcaacctg tctatcaagg acatcctgga gccggatgcc
     1081 gcggaaccag acgtgcagcg tggcaggagc ttccacttct acgatgccat ggatgggcag
     1141 atacagggca gcgtggagct ggcagcccca ggacaggcaa agatcgcagg cggggccgcg
     1201 gtgtctgaca gctccagcac ctcaatgaat gtgtactcgc tgagtgtgga ccctaacacc
     1261 tggcagactc tgctccatga gaggcacctg cggcagccag aacacaaagt cctgcagcag
     1321 ctgcgcagcc gcggggacaa cgtgtacgtg gtgactgagg tgctgcagac acagaaggag
     1381 gtggaagtca cgcgcaccca caagcgggag ggctcgggcc ggttttccct gcccggagcc
     1441 acgtgcttgc agggtgaggg ccagggccat ctgagccaga agaagacggt caccatcccc
     1501 tcaggcagca ccctcgcatt ccgggtggcc cagctggtta ttgactctga cttggacgtc
     1561 cttctcttcc cggataagaa gcagaggacc ttccagccac ccgcgacagg ccacaagcgt
     1621 tccacgagcg aaggcgcctg gccacagctg ccctctggcc tctccatgat gaggtgcctc
     1681 cacaacttcc tgacagatgg ggtccctgcg gagggggcgt tcactgaaga cttccagggc
     1741 ctacgggcag aggtggagac catctccaag gaactggagc ttttggacag agagctgtgc
     1801 cagctgctgc tggagggcct ggagggggtg ctgcgggacc agctggccct gcgagccttg
     1861 gaggaggcgc tggagcaggg ccagagcctt gggccggtgg agcccctgga cggtccagca
     1921 ggtgctgtcc tggagtgcct ggtgttgtcc tccggaatgc tggtgccgga actcgctatc
     1981 cctgttgtct acctgctggg ggcactgacc atgctgagtg aaacgcagca caagctgctg
     2041 gcggaggcgc tggagtcgca gaccctgttg gggccgctcg agctggtggg cagcctcttg
     2101 gagcagagtg ccccgtggca ggagcgcagc accatgtccc tgccccccgg gctcctgggg
     2161 aacagctggg gcgaaggagc accggcctgg gtcttgctgg acgagtgtgg cctagagctg
     2221 ggggaggaca ctccccacgt gtgctgggag ccgcaggccc agggccgcat gtgtgcactc
     2281 tacgcctccc tggcactgct atcaggactg agccaggagc cccactaccc ttatgacgtc
     2341 cccgattacg cctgagcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     2401 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2461 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2521 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2581 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2641 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2821 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2881 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3001 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3361 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3421 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3481 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3541 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3601 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3661 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3721 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3781 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3841 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3901 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3961 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4021 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4081 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4141 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4201 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4261 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4321 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4501 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4561 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6781 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
