LOCUS       V040238                 9691 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040238
VERSION     V040238
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..4545
                     /label="STAT4(NM_001243835)"
                     /note="STAT4(NM_001243835)"
                     /gene="STAT4"
     CDS             4552..4617
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4661..4872
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4885..5153
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5180..5779
                     /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    5786..6374
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6448..6681
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6753..6874
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6893..7028
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7066..7082)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7090..7106
                     /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(7114..7144)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7159..7180
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7468..8056)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8227..9087)
                     /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(9088..9192)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9666..9682
                     /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 catgtctcag tggaatcaag tccaacagtt agaaatcaag
     2341 tttttggagc aggtggatca attctatgat gacaactttc ccatggaaat tcggcatctg
     2401 ttggcccaat ggattgaaaa tcaagactgg gaggcagctt ctaacaatga aaccatggca
     2461 acgattcttc ttcaaaactt gttaatacaa ctggatgaac agttaggtcg tgtttccaaa
     2521 gagaaaaacc tactcttgat acacaatcta aaaagaatta ggaaggtcct tcagggaaaa
     2581 tttcatggaa atccaatgca tgtagctgtg gttatttcaa actgtttaag ggaagagagg
     2641 agaatattgg ctgcagccaa catgcctgtc caggggcctc tagagaaatc cttacaaagt
     2701 tcttcagttt cagaaagaca gaggaatgtg gagcacaaag tggctgccat taaaaacagt
     2761 gtgcagatga cagaacaaga taccaaatac ttagaagatc tgcaagacga atttgactac
     2821 aggtataaaa caattcagac aatggatcag agtgacaaga atagtgccat ggtgaatcag
     2881 gaagttttga cactgcagga aatgcttaac agcctcgatt tcaagagaaa ggaggctctc
     2941 agtaaaatga cccaaatcat ccatgagaca gacctgttaa tgaacaccat gctcatagaa
     3001 gagctgcaag actggaagcg gcggcagcaa atcgcctgca tcgggggtcc actccacaat
     3061 gggctcgacc agcttcagaa ctgctttaca ctattggcag aaagtctttt ccaactgaga
     3121 aggcaattgg agaaactaga ggagcaatct accaaaatga catatgaagg tgatcccatt
     3181 ccaatgcaaa gaactcacat gctagaaaga gtcaccttct tgatctacaa ccttttcaag
     3241 aactcatttg tggttgagcg acagccatgt atgccaaccc accctcagag gccgttggta
     3301 cttaaaaccc taattcagtt cactgtaaaa ctaaggctac taataaaatt gccagaacta
     3361 aactatcagg taaaggttaa ggcatcaatt gacaagaatg tttcaactct aagcaaccga
     3421 agatttgtac tttgtggaac taatgtcaaa gccatgtcta ttgaagaatc ttccaatggg
     3481 agtctctcag tagaatttcg acatttgcaa ccaaaggaaa tgaagtccag tgctggaggt
     3541 aaaggaaatg agggctgtca catggtgact gaagaacttc attccataac gtttgaaaca
     3601 cagatctgcc tctatggcct gaccatagat ttggagacca gctcattgcc tgtggtgatg
     3661 atttccaatg tcagtcagtt acctaatgct tgggcatcca tcatttggta caacgtgtca
     3721 accaacgatt cccagaactt ggttttcttt aataatcctc cacctgccac attgagtcaa
     3781 ctactggagg tgatgagctg gcagttttca tcgtacgttg gtcgtggtct taactcagat
     3841 caactccata tgctggcaga gaagcttaca gtccaatcta gctacagtga tggtcacctc
     3901 acctgggcca agttctgcaa ggaacattta cctggtaaat catttacctt ttggacatgg
     3961 cttgaagcaa tattggatct aattaagaaa cacattcttc ccctttggat tgatgggtat
     4021 gtcatgggct ttgttagcaa agagaaggaa cggctgttgc taaaggataa aatgcctggc
     4081 acctttttat taagattcag tgaaagccat ctcggaggaa taactttcac ctgggtggac
     4141 cattctgaaa gtggggaagt gagattccac tctgtagaac cctacaataa aggccggttg
     4201 tctgctctgc cattcgctga catcctgcga gactacaaag ttattatggc tgaaaacatt
     4261 cctgaaaacc ctctgaagta cctatatcct gacattccca aagacaaagc cttcggtaaa
     4321 cactacagct ctcagccttg cgaagtttca agaccaacag aaaggggtga caaaggttat
     4381 gttccttctg tttttatccc catctcaaca atccgaagtg attcaacaga gccacattct
     4441 ccatcagacc ttcttcccat gtctccaagt gtgtatgcgg tgttgagaga aaacctgagt
     4501 cccacaacaa ttgaaactgc aatgaagtct ccttattctg ctgaaggatc cgactacaaa
     4561 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     4621 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     4681 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4741 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4801 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4861 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4921 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     4981 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     5041 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     5101 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     5161 tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5221 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5281 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5341 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5401 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5461 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5521 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5581 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5641 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5701 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5761 ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5821 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5881 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5941 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     6001 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     6061 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6121 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6181 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6241 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6301 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6361 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6421 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6481 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6541 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6601 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6661 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6721 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6781 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6841 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6901 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6961 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7021 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     7081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9181 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9241 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9301 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9361 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9421 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9481 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9541 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9601 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9661 acgttgtaaa acgacggcca gtgccaagct g
//
