LOCUS       V024145                 9957 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024145
VERSION     V024145
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             2301..4802
                     /label="PROM2(NM_001165978)"
                     /note="PROM2(NM_001165978)"
                     /gene="PROM2"
     CDS             4818..4883
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4927..5138
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5151..5419
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5446..6045
                     /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    6052..6640
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6714..6947
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7019..7140
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7159..7294
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7332..7348)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7356..7372
                     /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(7380..7410)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7425..7446
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7734..8322)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8493..9353)
                     /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(9354..9458)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9932..9948
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgaagcaca cactggctct gctggctccc ctgctgggcc
     2341 tgggcctggg gctggccctg agtcagctgg ctgcaggggc cacagactgc aagttccttg
     2401 gcccggcaga gcacctgaca ttcaccccag cagccagggc ccggtggctg gcccctcgag
     2461 ttcgtgcgcc aggactcctg gactccctct atggcaccgt gcgccgcttc ctctcggtgg
     2521 tgcagctcaa tcctttccct tcagagttgg taaaggccct actgaatgag ctggcctccg
     2581 tgaaggtgaa tgaggtggtg cggtacgagg cgggctacgt ggtatgcgct gtgatcgcgg
     2641 gcctctacct gctgctggtg cccactgccg ggctttgctt ctgctgctgc cgctgccacc
     2701 ggcgctgcgg gggacgagtg aagacagagc acaaggcgct ggcctgtgag cgcgcggccc
     2761 tcatggtctt cctgctgctg accaccctct tgctgctgat tggtgtggtc tgtgcctttg
     2821 tcaccaacca gcgcacgcat gaacagatgg gccccagcat cgaggccatg cctgagaccc
     2881 tgctcagcct ctggggcctg gtctctgatg tcccccaaga gctgcaggcc gtggcacagc
     2941 aattctccct gccccaggag caagtctcag aggagctgga tggtgttggt gtgagcattg
     3001 ggagcgcgat ccacactcag ctcaggagct ccgtgtaccc cttgctggcg gccgtgggca
     3061 gtttgggcca ggtcctgcag gtctccgtgc accacctgca aaccttgaat gctacagtgg
     3121 tagagctgca ggccgggcag caggacctgg agccagccat ccgggaacac cgggaccgcc
     3181 tccttgagct gctgcaggag gccaggtgcc agggagattg tgcaggggcc ctgagctggg
     3241 cccgcaccct ggagctgggt gctgacttca gccaggtgcc ctctgtggac catgtcctgc
     3301 accagctaaa aggtgtcccc gaggccaact tctccagcat ggtccaggag gagaacagca
     3361 ccttcaacgc ccttccagcc ctggctgcca tgcagacatc cagcgtggtg caagagctga
     3421 agaaggcagt ggcccagcag ccggaagggg tgaggacact ggctgaaggg ttcccgggct
     3481 tggaggcagc ttcccgctgg gcccaggcac tgcaggaggt ggaggagagc agccgcccct
     3541 acctgcagga ggtgcagaga tacgagacct acaggtggat cgtgggctgc gtgctgtgct
     3601 ccgtggtcct attcgtggtg ctctgcaacc tgctgggcct caatctgggc atctggggcc
     3661 tgtctgccag ggacgacccc agccacccag aagccaaggg cgaggctgga gcccgcttcc
     3721 tcatggcagg tgtgggcctc agcttcctct ttgctgcacc cctcatcctc ctggtgttcg
     3781 ccaccttcct ggtgggtggc aacgtgcaga cgctggtgtg ccagagctgg gagaacggcg
     3841 agctctttga gtttgcagac accccaggga acctgccccc gtccatgaac ctgtcgcaac
     3901 ttcttggcct gaggaagaac atcagcatcc accaagccta tcagcagtgc aaggaagggg
     3961 cagcgctctg gacagtcctg cagctcaacg actcctacga cctggaggag cacctggata
     4021 tcaaccagta taccaacaag ctacggcagg agttgcagag cctgaaagta gacacacaga
     4081 gcctggacct gctgagctca gccgcccgcc gggacctgga ggccctgcag agcagtgggc
     4141 ttcagcgcat ccactacccc gacttcctcg ttcagatcca gaggcccgtg gtgaagacca
     4201 gcatggagca gctggcccag gagctgcaag gactggccca ggcccaagac aattctgtgc
     4261 tggggcagcg gctgcaggag gaggcccaag gactcagaaa ccttcaccag gagaaggtcg
     4321 tcccccagca gagccttgtg gcaaagctca acctcagcgt cagggccctg gagtcctctg
     4381 ccccgaatct ccagctggag acctcagatg tcctagccaa tgtcacctac ctgaaaggag
     4441 agctgcctgc ctgggcagcc aggatcctga ggaatgtgag tgagtgtttc ctggcccggg
     4501 agatgggcta cttctcccag tacgtggcct gggtgagaga ggaggtgact cagcgcattg
     4561 ccacctgcca gcccctctcc ggagccctgg acaacagccg tgtgatcctg tgtgacatga
     4621 tggctgaccc ctggaatgcc ttctggttct gcctggcatg gtgcaccttc ttcctgatcc
     4681 ccagcatcat ctttgccgtc aagacctcca aatacttccg tcctatccgg aaacgcctca
     4741 gctccaccag ctctgaggag actcagctct tccacatccc ccgggttacc tccctgaagc
     4801 tgggtggagg cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     4861 actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     4921 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     4981 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     5041 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     5101 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     5161 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     5221 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     5281 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     5341 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     5401 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag
     5461 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     5521 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     5581 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5641 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5701 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5761 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5821 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5881 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5941 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     6001 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct
     6061 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     6121 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     6181 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     6241 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     6301 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     6361 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     6421 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     6481 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     6541 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     6601 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     6661 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6721 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     6781 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6841 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6901 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6961 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     7021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7141 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     7201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     7261 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     7321 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7381 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7441 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7501 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7561 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7621 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8161 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8401 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8461 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8521 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8581 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8641 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8701 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8761 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8821 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8881 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8941 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     9001 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     9061 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     9121 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9181 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9241 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9301 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9361 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9421 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9481 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     9541 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     9601 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     9661 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     9721 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     9781 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     9841 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9901 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
