LOCUS       V025311                 8877 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025311
VERSION     V025311
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..3731
                     /label="TRIM72(NM_001008274)"
                     /note="TRIM72(NM_001008274)"
                     /gene="TRIM72"
     CDS             3738..3803
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3847..4058
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4071..4339
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4366..4965
                     /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    4972..5560
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5634..5867
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5939..6060
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6079..6214
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6252..6268)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6276..6292
                     /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(6300..6330)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6345..6366
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6654..7242)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7413..8273)
                     /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(8274..8378)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8852..8868
                     /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 atgtcggctg cgcccggcct cctgcaccag gagctgtcct
     2341 gcccgctgtg cctgcagctg ttcgacgcgc ccgtgacagc cgagtgcggc cacagtttct
     2401 gccgcgcctg cctaggccgc gtggccgggg agccggcggc ggatggcacc gttctctgcc
     2461 cctgctgcca ggcccccacg cggccgcagg cactcagcac caacctgcag ctggcgcgcc
     2521 tggtggaggg gctggcccag gtgccgcagg gccactgcga ggagcacctg gacccgctga
     2581 gcatctactg cgagcaggac cgcgcgctgg tgtgcggagt gtgcgcctca ctcggctcgc
     2641 accgcggtca tcgcctcctg cctgccgccg aggcccacgc acgcctcaag acacagctgc
     2701 cacagcagaa actgcagctg caggaggcat gcatgcgcaa ggagaagagt gtggctgtgc
     2761 tggagcatca gctggtggag gtggaggaga cagtgcgtca gttccggggg gccgtggggg
     2821 agcagctggg caagatgcgg gtgttcctgg ctgcactgga gggctccttg gaccgcgagg
     2881 cagagcgtgt acggggtgag gcaggggtcg ccttgcgccg ggagctgggg agcctgaact
     2941 cttacctgga gcagctgcgg cagatggaga aggtcctgga ggaggtggcg gacaagccgc
     3001 agactgagtt cctcatgaaa tactgcctgg tgaccagcag gctgcagaag atcctggcag
     3061 agtctccccc acccgcccgt ctggacatcc agctgccaat tatctcagat gacttcaaat
     3121 tccaggtgtg gaggaagatg ttccgggctc tgatgccagc gctggaggag ctgacctttg
     3181 acccgagctc tgcgcacccg agcctggtgg tgtcttcctc tggccgccgc gtggagtgct
     3241 cggagcagaa ggcgccgccg gccggggagg acccgcgcca gttcgacaag gcggtggcgg
     3301 tggtggcgca ccagcagctc tccgagggcg agcactactg ggaggtggat gttggcgaca
     3361 agccgcgctg ggcgctgggc gtgatcgcgg ccgaggcccc ccgccgcggg cgcctgcacg
     3421 cggtgccctc gcagggcctg tggctgctgg ggctgcgcga gggcaagatc ctggaggcac
     3481 acgtggaggc caaggagccg cgcgctctgc gcagccccga gaggcggccc acgcgcattg
     3541 gcctttacct gagcttcggc gacggcgtcc tctccttcta cgatgccagc gacgccgacg
     3601 cgctcgtgcc gctttttgcc ttccacgagc gcctgcccag gcccgtgtac cccttcttcg
     3661 acgtgtgctg gcacgacaag ggcaagaatg cccagccgct gctgctcgtg ggtcccgaag
     3721 gcgccgaggc cggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     3781 actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     3841 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3901 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3961 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     4021 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     4081 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     4141 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     4201 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     4261 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     4321 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag
     4381 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     4441 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     4501 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4561 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4621 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4681 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4741 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4801 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4861 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4921 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct
     4981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5341 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     5401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5521 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     5581 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5641 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     5701 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5761 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5821 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5881 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6061 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     6121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6181 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     6241 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6301 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6361 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6421 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6481 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6541 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7081 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7261 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7321 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7381 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7441 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7501 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7561 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7621 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7681 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7741 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7801 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7861 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7921 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7981 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8041 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8101 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8161 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8221 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8281 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8341 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8401 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8461 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     8521 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     8581 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     8641 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     8701 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8761 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8821 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
