LOCUS       V032700                 9037 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032700
VERSION     V032700
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..3891
                     /label="PSIP1(NM_033222)"
                     /note="PSIP1(NM_033222)"
                     /gene="PSIP1"
     CDS             3898..3963
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4007..4218
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4231..4499
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4526..5125
                     /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    5132..5720
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5794..6027
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6099..6220
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6239..6374
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6412..6428)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6436..6452
                     /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(6460..6490)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6505..6526
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6814..7402)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7573..8433)
                     /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(8434..8538)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9012..9028
                     /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 catgactcgc gatttcaaac ctggagacct catcttcgcc
     2341 aagatgaaag gttatcccca ttggccagct cgagtagacg aagttcctga tggagctgta
     2401 aagccaccca caaacaaact acccattttc ttttttggaa ctcatgagac tgctttttta
     2461 ggaccaaagg atatatttcc ttactcagaa aataaggaaa agtatggcaa accaaataaa
     2521 agaaaaggtt ttaatgaagg tttatgggag atagataaca atccaaaagt gaaattttca
     2581 agtcaacagg cagcaactaa acaatcaaat gcatcatctg atgttgaagt tgaagaaaag
     2641 gaaactagtg tttcaaagga agataccgac catgaagaaa aagccagcaa tgaggatgtg
     2701 actaaagcag ttgacataac tactccaaaa gctgccagaa gggggagaaa gagaaaggca
     2761 gaaaaacaag tagaaactga ggaggcagga gtagtcacaa cagcaacagc atctgttaat
     2821 ctaaaagtga gtcctaaaag aggacgacct gcagctacag aagtcaagat tccaaaacca
     2881 agaggcagac ccaaaatggt aaaacagccc tgtccttcag agagtgacat cattactgaa
     2941 gaggacaaaa gtaagaaaaa ggggcaagag gaaaaacaac ctaaaaagca gcctaagaag
     3001 gatgaagagg gccagaagga agaagataag ccaagaaaag agccggataa aaaagagggg
     3061 aagaaagaag ttgaatcaaa aaggaaaaat ttagctaaaa caggggttac ttcaacctcc
     3121 gattctgaag aagaaggaga tgatcaagaa ggtgaaaaga agagaaaagg tgggaggaac
     3181 tttcagactg ctcacagaag gaatatgctg aaaggccaac atgagaaaga agcagcagat
     3241 cgaaaacgca agcaagagga acaaatggaa actgagcagc agaataaaga tgaaggaaag
     3301 aagccagaag ttaagaaagt ggagaagaag cgagaaacat caatggattc tcgacttcaa
     3361 aggatacatg ctgagattaa aaattcactc aaaattgata atcttgatgt gaacagatgc
     3421 attgaggcct tggatgaact tgcttcactt caggtcacaa tgcaacaagc tcagaaacac
     3481 acagagatga ttactacact gaaaaaaata cggcgattca aagttagtca ggtaatcatg
     3541 gaaaagtcta caatgttgta taacaagttt aagaacatgt tcttggttgg tgaaggagat
     3601 tccgtgatca cccaagtgct gaataaatct cttgctgaac aaagacagca tgaggaagcg
     3661 aataaaacca aagatcaagg gaagaaaggg ccaaacaaaa agctagagaa ggaacaaaca
     3721 gggtcaaaga ctctaaatgg aggatctgat gctcaagatg gtaatcagcc acaacataac
     3781 ggggagagca atgaagacag caaagacaac catgaagcca gcacgaagaa aaagccatcc
     3841 agtgaagaga gagagactga aatatctctg aaggattcta cactagataa cggatccgac
     3901 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3961 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     4021 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     4081 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     4141 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     4201 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     4261 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     4321 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4381 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4441 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4501 aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc gcctcgccac
     4561 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4621 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4681 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4741 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4801 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4861 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4921 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4981 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5041 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5101 catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca aaatttgtga
     5161 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5221 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5281 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5341 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5401 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5461 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5521 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5581 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5641 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5701 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     5761 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5821 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5881 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5941 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6001 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6061 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6121 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6181 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6241 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6301 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6361 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6421 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6481 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6541 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6601 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8581 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     8641 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     8701 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     8761 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     8821 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     8881 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     8941 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9001 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
