LOCUS       V036928                 9067 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036928
VERSION     V036928
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2305..2394
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2401..3978
                     /label="TBC1D19(NM_018317)"
                     /note="TBC1D19(NM_018317)"
                     /gene="TBC1D19"
     promoter        4037..4248
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4261..4529
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4556..5155
                     /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    5162..5750
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5824..6057
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6129..6250
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6269..6404
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6442..6458)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6466..6482
                     /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(6490..6520)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6535..6556
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6844..7432)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7603..8463)
                     /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(8464..8568)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9042..9058
                     /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 catggaacaa aaactaatat cagaggaaga tttggaacag
     2341 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttactcgag
     2401 atgttgcagg aggagtcgga cctctctctc attattgccc agatagtcca aaagctcaag
     2461 ggctccaatt tgtactctca gctggaacgg caggcctggg ccagtcttca gagacctgag
     2521 attaaacttg aatcactgaa agaagatatt aaggaattct ttaaaatatc aggttgggag
     2581 aagaaacttc agaatgctgt ttatagtgaa ctgagtgtgt ttcctttacc tagtcatcct
     2641 gctgcacctc ctgaacatct taaagaacct ttggtataca tgaggaaagc acagggaagt
     2701 tgggaaaaaa gaattttgaa gagtttaaat agtatgtgca ctgaactgag tatcccactg
     2761 gcacgaaaga ggccagttgg agaacagaaa gaacttctta ataaatggaa tgaaatggga
     2821 actgatgaac cagatttaag ccttttcaga cctgtttatg cacctaagga ttttcttgag
     2881 gtattaatta atcttcgcaa cccaaattat gaaaacggtg attctcttag tttcaggact
     2941 catttgggtt taattcaagt tccactgaaa gtaaaagaca tccctgaatt gaaagaatgc
     3001 tttgtggaac ttggcttaaa tataggacaa ctgggtatag atgattctac acaagtgcct
     3061 cctgaacttt ttgaaaatga acatgtacgt attgggcaaa aagttctagc agaacaagat
     3121 agtgctgctg ctcaacagta catcagacaa ggaagtccca cggcactgag agctgaattg
     3181 tgggctctca ttttgaatat ttccagccaa cctgaggatg tcttgtatta tgagcagctt
     3241 aagaccaatg tgatacaaca tgaccttttg gtggacagtc taatctataa agatgtgaag
     3301 ttgacagcaa gcaatgatga ttattatttt gtatttgaag attatttata tcaggtatta
     3361 ctttgttttt cccgggatac atctgtgttg agtcactttg cattcaacag tgcctcgcca
     3421 ccaaaatcat acataagagg aaaactagga ctggaagaat atgctgtctt ttacccacca
     3481 aatggtgtta tcccttttca tggattttca atgtatgttg caccactttg ttttctatac
     3541 catgaacctt ccaaattgta tcagatattc cgtgagatgt atgtgcgttt tttcttcaga
     3601 ctccattcca tctcttctca tccttctggt attgtgtcac tctgtctgct gtttgaaact
     3661 cttcttcaaa cttatcttcc ccaactcttt tatcatctac gagaaattgg ggctcaacca
     3721 cttcgcatat catttaagtg gatggttcga gctttctctg gatacttagc tacagatcag
     3781 ctcttgcttt tatgggatag aatcctagga tacaactctc tggaaattct tgctgtgctg
     3841 gcagctgccg tgtttgcttt ccgagcagtg aacctgatgg aggtgacatc actggctgca
     3901 gctgaagcag ttcttgctga cctttctact ttaaaagtta tgcctcttct tcagattttt
     3961 ctgtttgcta ctgtcacctg aggatccgat atctagcggc cgcgaaggat ctgcgatcgc
     4021 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     4081 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     4141 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     4201 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     4261 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     4321 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     4381 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     4441 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     4501 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac
     4561 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     4621 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     4681 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4741 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4801 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4861 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4921 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4981 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5041 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5101 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga
     5161 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5221 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5281 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5341 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5401 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5461 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5521 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5581 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5641 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5701 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
     5761 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5821 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     5881 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5941 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6001 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6061 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6121 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6241 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6421 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7261 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8581 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8641 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8701 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8761 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     8821 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     8881 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8941 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9001 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9061 caagctg
//
