LOCUS       V033620                 8782 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033620
VERSION     V033620
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3636
                     /label="GSDMA(NM_178171)"
                     /note="GSDMA(NM_178171)"
                     /gene="GSDMA"
     CDS             3643..3708
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3752..3963
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3976..4244
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4271..4870
                     /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    4877..5465
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5539..5772
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5844..5965
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5984..6119
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6157..6173)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6181..6197
                     /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(6205..6235)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6250..6271
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6559..7147)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7318..8178)
                     /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(8179..8283)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8757..8773
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgaccatg tttgaaaatg tcacccgggc cctggccaga
     2341 cagctaaacc ctcgagggga cctgacacca cttgacagcc tcatcgactt caagcgcttc
     2401 catcccttct gcctggtgct gaggaagagg aagagcacgc tcttctgggg ggcccggtac
     2461 gtccgcaccg actacacgct gctggatgtg cttgagcccg gcagctcacc ttcagaccca
     2521 acagacactg ggaattttgg ctttaagaat atgctggaca cccgagtgga gggagatgtg
     2581 gatgtaccaa agacggtgaa ggtgaaggga acggcagggc tctcgcagaa cagcactctg
     2641 gaggtccaga cactcagtgt ggctcccaag gccctggaga ccgtgcagga gaggaagctg
     2701 gcagcagacc acccattcct gaaggagatg caagatcaag gggagaacct gtatgtggtg
     2761 atggaggtgg tggagacggt gcaggaggtc acactggagc gagccggcaa ggcagaggcc
     2821 tgcttctccc tccccttctt cgccccattg gggctacagg gatccataaa tcacaaggag
     2881 gctgtaacca tccccaaggg ctgcgtcctg gcctttcgag tgagacagct gatggtcaaa
     2941 ggcaaagatg agtgggatat tccacatatc tgcaatgata acatgcaaac cttccctcct
     3001 ggagaaaagt caggagagga gaaggtcatc cttatccagg catctgatgt tggggacgta
     3061 cacgaaggct tcaggacact aaaagaagaa gttcagagag agacccaaca agtggagaag
     3121 ctgagccgag tagggcaaag ctccctgctc agctccctca gcaaacttct agggaagaaa
     3181 aaggagctac aagaccttga gctcgcactt gaaggggctc tagacaaggg acatgaagtg
     3241 accctggagg cactcccaaa agatgtcctg ctatcaaagg aggccgtggg cgccatcctc
     3301 tatttcgttg gagccctaac agagctaagt gaagcccaac agaagctgct ggtgaaatcc
     3361 atggagaaaa agatcctacc cgtgcagcta aagctggtgg agagcacgat ggaacagaac
     3421 ttcctgctgg ataaagaggg tgttttcccc ctgcaacctg agctgctctc ctcccttggg
     3481 gacgaggagc tgaccctcac ggaggctcta gtcgggctga gtggcctgga agtgcagaga
     3541 tcgggccccc aatatatgtg ggacccagac accctccctc gcctctgtgc tctttatgca
     3601 ggcctctctc tccttcagca gcttaccaag gcctccggat ccgactacaa agaccatgac
     3661 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga
     3721 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3781 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     3841 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3901 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3961 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     4021 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4081 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4141 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4201 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4261 cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     4321 ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     4381 cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     4441 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     4501 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4561 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4621 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4681 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4741 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4801 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4861 cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4921 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4981 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5041 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5101 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5161 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5221 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5281 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5341 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5401 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5461 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5521 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     5581 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5641 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     5701 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     5761 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     5821 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     5881 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5941 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6001 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6061 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6121 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     6181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6961 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8281 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8341 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     8401 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     8461 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     8521 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     8581 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     8641 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     8701 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     8761 aacgacggcc agtgccaagc tg
//
