LOCUS       V020943                10215 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020943
VERSION     V020943
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2807..3523
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Aequorea coerulescens GFP"
                     /transl_table=1
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
                     /label="AcGFP1"
     CDS             3557..5008
                     /label="GSDMD(NM_024736)"
                     /note="GSDMD(NM_024736)"
                     /gene="GSDMD"
     promoter        5050..5549
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5570..6169
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    6183..6771
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6978..7611
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7740..7756)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7764..7780
                     /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(7788..7818)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7833..7854
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8142..8730)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8901..9761)
                     /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(9762..9866)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9914..10048
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctaccggtc gccaccatgg tgagcaaggg
     2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg
     3541 aggaggaggg ggaagcatgg ggtcggcctt tgagcgggta gtccggagag tggtccagga
     3601 gctggaccat ggtggggagt tcatccctgt gaccagcctg cagagctcca ctggcttcca
     3661 gccctactgc ctggtggtta ggaagccctc aagctcatgg ttctggaaac cccgttataa
     3721 gtgtgtcaac ctgtctatca aggacatcct ggagccggat gccgcggaac cagacgtgca
     3781 gcgtggcagg agcttccact tctacgatgc catggatggg cagatacagg gcagcgtgga
     3841 gctggcagcc ccaggacagg caaagatcgc aggcggggcc gcggtgtctg acagctccag
     3901 cacctcaatg aatgtgtact cgctgagtgt ggaccctaac acctggcaga ctctgctcca
     3961 tgagaggcac ctgcggcagc cagaacacaa agtcctgcag cagctgcgca gccgcgggga
     4021 caacgtgtac gtggtgactg aggtgctgca gacacagaag gaggtggaag tcacgcgcac
     4081 ccacaagcgg gagggctcgg gccggttttc cctgcccgga gccacgtgct tgcagggtga
     4141 gggccagggc catctgagcc agaagaagac ggtcaccatc ccctcaggca gcaccctcgc
     4201 attccgggtg gcccagctgg ttattgactc tgacttggac gtccttctct tcccggataa
     4261 gaagcagagg accttccagc cacccgcgac aggccacaag cgttccacga gcgaaggcgc
     4321 ctggccacag ctgccctctg gcctctccat gatgaggtgc ctccacaact tcctgacaga
     4381 tggggtccct gcggaggggg cgttcactga agacttccag ggcctacggg cagaggtgga
     4441 gaccatctcc aaggaactgg agcttttgga cagagagctg tgccagctgc tgctggaggg
     4501 cctggagggg gtgctgcggg accagctggc cctgcgagcc ttggaggagg cgctggagca
     4561 gggccagagc cttgggccgg tggagcccct ggacggtcca gcaggtgctg tcctggagtg
     4621 cctggtgttg tcctccggaa tgctggtgcc ggaactcgct atccctgttg tctacctgct
     4681 gggggcactg accatgctga gtgaaacgca gcacaagctg ctggcggagg cgctggagtc
     4741 gcagaccctg ttggggccgc tcgagctggt gggcagcctc ttggagcaga gtgccccgtg
     4801 gcaggagcgc agcaccatgt ccctgccccc cgggctcctg gggaacagct ggggcgaagg
     4861 agcaccggcc tgggtcttgc tggacgagtg tggcctagag ctgggggagg acactcccca
     4921 cgtgtgctgg gagccgcagg cccagggccg catgtgtgca ctctacgcct ccctggcact
     4981 gctatcagga ctgagccagg agccccacta gggatccacc ggatctagat aactgatcat
     5041 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     5101 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     5161 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     5221 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     5281 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     5341 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     5401 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     5461 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     5521 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta
     5581 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     5641 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     5701 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5761 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5821 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5881 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5941 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     6001 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     6061 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     6121 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga
     6181 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     6241 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     6301 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     6361 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     6421 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     6481 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6541 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     6601 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6661 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6721 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
     6781 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     6841 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     6901 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6961 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     7021 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     7081 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     7141 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     7201 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     7261 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     7321 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     7381 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     7441 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7501 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7561 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7621 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7681 ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg
     7741 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7801 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7861 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7921 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7981 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8581 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8821 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8881 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8941 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9001 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9061 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     9901 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     9961 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    10021 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    10081 caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta
    10141 aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac
    10201 tacaaactta gtagt
//
