LOCUS       V031660                 8857 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031660
VERSION     V031660
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..3711
                     /label="TRAF4(NM_004295)"
                     /note="TRAF4(NM_004295)"
                     /gene="TRAF4"
     CDS             3718..3783
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3827..4038
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4051..4319
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4346..4945
                     /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    4952..5540
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5614..5847
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5919..6040
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6059..6194
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6232..6248)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6256..6272
                     /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(6280..6310)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6325..6346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6634..7222)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7393..8253)
                     /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(8254..8358)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8832..8848
                     /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 catgcctggc ttcgactaca agttcctgga gaagcccaag
     2341 cgacggctgc tgtgcccact gtgcgggaag cccatgcgcg agcctgtgca ggtttccacc
     2401 tgcggccacc gtttctgcga tacctgcctg caggagttcc tcagtgaagg agtcttcaag
     2461 tgccctgagg accagcttcc tctggactat gccaagatct acccagaccc ggagctggaa
     2521 gtacaagtat tgggcctgcc tatccgctgc atccacagtg aggagggctg ccgctggagt
     2581 gggccactac gtcatctaca gggccacctg aatacctgca gcttcaatgt cattccctgc
     2641 cctaatcgct gccccatgaa gctgagccgc cgtgatctac ctgcacactt gcagcatgac
     2701 tgccccaagc ggcgcctcaa gtgcgagttt tgtggctgtg acttcagtgg ggaggcctat
     2761 gagagccatg agggtatgtg cccccaggag agtgtctact gtgagaataa gtgtggtgcc
     2821 cgcatgatgc ggcggctgct ggcccagcat gccacctctg agtgccccaa gcgcactcag
     2881 ccctgcacct actgcactaa ggagttcgtc tttgacacca tccagagcca ccagtaccag
     2941 tgcccaaggc tgcctgttgc ctgccccaac caatgtggtg tgggcactgt ggctcgggag
     3001 gacctgccag gccatctgaa ggacagctgt aacaccgccc tggtgctctg cccattcaaa
     3061 gactccggct gcaagcacag gtgccctaag ctggcaatgg cacggcatgt ggaggagagt
     3121 gtgaagccac atctggccat gatgtgtgcc ctggtgagcc ggcaacggca ggagctgcag
     3181 gagcttcggc gagagctgga ggagctatca gtgggcagtg atggcgtgct catctggaag
     3241 attggcagct atggacggcg gctacaggag gccaaggcca agcccaacct tgagtgcttc
     3301 agcccagcct tctacacaca taagtatggt tacaagctgc aggtgtctgc attcctcaat
     3361 ggcaatggca gtggtgaggg cacacacctc tcactgtaca ttcgtgtgct gcctggtgcc
     3421 tttgacaatc tccttgagtg gccctttgcc cgccgtgtca ccttctccct gctggatcag
     3481 agcgaccctg ggctggctaa accacagcac gtcactgaga ccttccaccc cgacccaaac
     3541 tggaagaatt tccagaagcc aggcacgtgg cggggctccc tggatgagag ttctctgggc
     3601 tttggttatc ccaagttcat ctcccaccag gacattcgaa agcgaaacta tgtgcgggat
     3661 gatgcagtct tcatccgtgc tgctgttgaa ctgccccgga agatcctcag cggatccgac
     3721 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3781 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3841 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     3901 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3961 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4021 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     4081 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     4141 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4201 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4261 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4321 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     4381 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4441 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4501 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4561 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4621 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4681 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4741 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4801 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4861 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4921 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     4981 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5041 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5101 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5161 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5221 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5281 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5341 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5401 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5461 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5521 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     5581 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5641 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5701 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5761 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5821 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     5881 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     5941 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6001 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6061 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6121 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6181 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6241 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7141 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7201 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7261 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7321 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7381 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7441 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7501 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7561 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7621 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7681 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7741 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7801 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7861 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7921 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7981 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8041 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8101 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8161 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8221 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8281 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8341 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8401 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     8461 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     8521 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     8581 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     8641 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     8701 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     8761 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     8821 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
