LOCUS       V021238                 9640 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021238
VERSION     V021238
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3754
                     /label="GSDMD(NM_024736)"
                     /note="GSDMD(NM_024736)"
                     /gene="GSDMD"
     promoter        3799..4010
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4023..4291
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4320..4329
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4350..5012
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             5082..5135
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             5136..5735
                     /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    5736..6323
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6397..6630
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6702..6823
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6842..6977
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7015..7031)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7039..7055
                     /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(7063..7093)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7108..7129
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7417..8005)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8176..9036)
                     /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(9037..9141)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9615..9631
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggggtc ggcctttgag cgggtagtcc ggagagtggt
     2341 ccaggagctg gaccatggtg gggagttcat ccctgtgacc agcctgcaga gctccactgg
     2401 cttccagccc tactgcctgg tggttaggaa gccctcaagc tcatggttct ggaaaccccg
     2461 ttataagtgt gtcaacctgt ctatcaagga catcctggag ccggatgccg cggaaccaga
     2521 cgtgcagcgt ggcaggagct tccacttcta cgatgccatg gatgggcaga tacagggcag
     2581 cgtggagctg gcagccccag gacaggcaaa gatcgcaggc ggggccgcgg tgtctgacag
     2641 ctccagcacc tcaatgaatg tgtactcgct gagtgtggac cctaacacct ggcagactct
     2701 gctccatgag aggcacctgc ggcagccaga acacaaagtc ctgcagcagc tgcgcagccg
     2761 cggggacaac gtgtacgtgg tgactgaggt gctgcagaca cagaaggagg tggaagtcac
     2821 gcgcacccac aagcgggagg gctcgggccg gttttccctg cccggagcca cgtgcttgca
     2881 gggtgagggc cagggccatc tgagccagaa gaagacggtc accatcccct caggcagcac
     2941 cctcgcattc cgggtggccc agctggttat tgactctgac ttggacgtcc ttctcttccc
     3001 ggataagaag cagaggacct tccagccacc cgcgacaggc cacaagcgtt ccacgagcga
     3061 aggcgcctgg ccacagctgc cctctggcct ctccatgatg aggtgcctcc acaacttcct
     3121 gacagatggg gtccctgcgg agggggcgtt cactgaagac ttccagggcc tacgggcaga
     3181 ggtggagacc atctccaagg aactggagct tttggacaga gagctgtgcc agctgctgct
     3241 ggagggcctg gagggggtgc tgcgggacca gctggccctg cgagccttgg aggaggcgct
     3301 ggagcagggc cagagccttg ggccggtgga gcccctggac ggtccagcag gtgctgtcct
     3361 ggagtgcctg gtgttgtcct ccggaatgct ggtgccggaa ctcgctatcc ctgttgtcta
     3421 cctgctgggg gcactgacca tgctgagtga aacgcagcac aagctgctgg cggaggcgct
     3481 ggagtcgcag accctgttgg ggccgctcga gctggtgggc agcctcttgg agcagagtgc
     3541 cccgtggcag gagcgcagca ccatgtccct gccccccggg ctcctgggga acagctgggg
     3601 cgaaggagca ccggcctggg tcttgctgga cgagtgtggc ctagagctgg gggaggacac
     3661 tccccacgtg tgctgggagc cgcaggccca gggccgcatg tgtgcactct acgcctccct
     3721 ggcactgcta tcaggactga gccaggagcc ccactaggcg gccgcgaagg atctgcgatc
     3781 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3841 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3901 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3961 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     4021 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     4081 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     4141 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     4201 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     4261 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg
     4321 ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg
     4381 gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg
     4441 gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc
     4501 tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga
     4561 accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg
     4621 aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg
     4681 gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga
     4741 tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg
     4801 gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca
     4861 gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt
     4921 tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc
     4981 agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg
     5041 ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc
     5101 taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc
     5161 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     5221 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     5281 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5341 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5401 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5461 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5521 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5581 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5641 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5701 gcacctggtg catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt
     5761 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5821 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5881 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5941 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     6001 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6061 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6121 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     6181 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6241 gcctgctgcc ggctctgcgg cctcttccgc gtctccgcct tcgccctcag acgagtcgga
     6301 tctccctttg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6361 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6421 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6481 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6541 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6601 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6661 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6721 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6781 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6841 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6901 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6961 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     7021 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7081 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9181 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9241 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9301 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9361 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9421 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9481 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9541 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9601 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
