LOCUS       V043728                 9937 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043728
VERSION     V043728
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..4857
                     /label="PIWIL4(NM_152431)"
                     /note="PIWIL4(NM_152431)"
                     /gene="PIWIL4"
     promoter        4907..5118
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5131..5399
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5426..6025
                     /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    6032..6620
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6694..6927
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6999..7120
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7139..7274
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7312..7328)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7336..7352
                     /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(7360..7390)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7405..7426
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7714..8302)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8473..9333)
                     /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(9334..9438)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9912..9928
                     /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 catgagtgga agagcccgag tgaaggccag aggcatcgcc
     2341 cgcagcccca gtgccacaga agtggggcgc atccaagcct cgccattgcc tagatctgtt
     2401 gatcttagta acaatgaagc atcctctagc aatggcttct tgggaacaag caggatctca
     2461 accaacgata aatatgggat atcttctggt gatgctggaa gtaccttcat ggaaagaggt
     2521 gtgaaaaaca aacaggactt tatggatttg agtatctgta ccagagaaaa attggcacat
     2581 gtgagaaatt gtaaaacagg ttccagtgga atacctgtga aactggttac aaacctcttt
     2641 aacttagatt ttccccaaga ctggcagcta taccagtacc atgtgacata tattccagat
     2701 ttagcatcta gaaggctgag aattgcttta ctttatagtc atagtgaact ttccaacaaa
     2761 gcaaaagcat tcgacggtgc catccttttt ctgtcacaaa agctagaaga aaaggtcaca
     2821 gagttgtcaa gtgaaactca aagaggtgag actataaaga tgactatcac cctgaagagg
     2881 gagctgccat caagttctcc cgtgtgcatc caggtcttca atatcatctt cagaaagatc
     2941 ctcaaaaagt tgtccatgta ccaaattgga cggaacttct ataatccttc agagccaatg
     3001 gaaattcccc agcacaaatt atccctttgg cctgggtttg ccatttctgt gtcatatttt
     3061 gaaaggaagc tcctgtttag tgctgatgtg agttacaaag tcctccggaa tgagacggtt
     3121 ctggaattca tgactgctct ctgtcaaaga actggcttgt cctgtttcac ccagacgtgt
     3181 gagaagcagc taatagggct cattgtcctt acaagataca ataacagaac ctactccatt
     3241 gatgacattg actggtcagt gaagcccaca cacacctttc agaagcggga tggcaccgag
     3301 atcacctatg tggattacta caagcagcag tatgatatta ctgtatcgga cctgaatcag
     3361 cccatgcttg ttagtctgtt aaagaagaag agaaatgaca acagtgaggc tcagctcgcc
     3421 cacctgatac ctgagctctg ctttctaaca gggctgactg accaggcaac atctgatttc
     3481 cagctgatga aggctgtggc tgaaaagaca cgtctcagtc cttcaggccg gcagcagcgc
     3541 ctggccaggc ttgtggacaa catccagagg aataccaatg ctcgctttga actagagacc
     3601 tggggactgc attttggaag ccagatatct ctgactggcc ggattgtgcc ttcagaaaaa
     3661 atattaatgc aagaccacat atgtcaacct gtgtctgctg ctgactggtc caaggatatt
     3721 cgaacttgca agattttaaa tgcacagtct ttgaatacct ggttgatttt atgtagcgac
     3781 agaactgaat atgttgccga gagctttctg aactgcttga gaagagttgc aggttccatg
     3841 ggatttaatg tggactaccc caaaatcata aaagtacaag aaaatccagc tgcatttgtt
     3901 agagctatac agcaatatgt tgatcctgat gttcagctgg taatgtgcat tctgccttct
     3961 aatcagaaga cctattatga ttccattaaa aaatatttga gctcagactg cccagtccca
     4021 agccaatgtg tgcttgctcg gaccttgaat aaacagggca tgatgatgag tatcgccacc
     4081 aagatcgcta tgcagatgac ttgcaagctc ggaggcgagc tgtgggctgt ggaaatacct
     4141 ttaaagtccc tgatggtggt cggtattgat gtctgtaaag atgcactcag caaggacgtg
     4201 atggttgttg gatgcgtggc cagtgttaac cccagaatca ccaggtggtt ttcccgctgt
     4261 atccttcaga gaacaatgac tgatgttgca gattgcttga aagttttcat gactggagca
     4321 ctcaacaaat ggtacaagta caatcatgat ttgccagcac ggataattgt gtaccgtgct
     4381 ggtgtagggg atggtcagct gaaaacactt attgaatatg aagtcccaca gctgctgagc
     4441 agtgtggcag aatccagctc aaataccagc tcaagactgt cggtgattgt ggtcaggaag
     4501 aagtgcatgc cacgattctt taccgaaatg aaccgcactg tacagaaccc cccacttggc
     4561 actgttgtgg attcagaagc aacacgtaac gaatggtatg acttttatct gatcagccag
     4621 gtggcctgcc ggggaactgt tagtcctacc tactataatg tcatctatga tgacaacggc
     4681 ttgaagcccg accatatgca gagacttaca ttcaaattgt gccacctgta ctacaactgg
     4741 ccgggcatag tcagtgtccc agcaccatgt cagtatgctc acaagctgac ctttctggtg
     4801 gcacaaagca ttcataaaga acccagtctg gaattagcca accatctctt ctacctgtag
     4861 gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4921 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4981 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     5041 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     5101 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     5161 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     5221 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     5281 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     5341 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     5401 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     5461 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5521 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5581 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5641 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5701 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5761 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5821 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5881 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5941 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     6001 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     6061 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6121 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6181 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6241 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6301 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6361 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6421 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6481 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6541 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6601 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6661 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6721 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6781 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6841 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6901 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6961 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     7021 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7081 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     7141 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7201 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7261 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     7321 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7381 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7501 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7561 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7621 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     8161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8521 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8581 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8641 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8701 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8761 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8821 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8881 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8941 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9001 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9061 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9121 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9181 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9241 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9301 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9361 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9421 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9481 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9541 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9601 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9661 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9721 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9781 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9841 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9901 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
