LOCUS       V021887                 8817 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021887
VERSION     V021887
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..3725
                     /label="TRIM21(NM_003141)"
                     /note="TRIM21(NM_003141)"
                     /gene="TRIM21"
     CDS             3726..3743
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3787..3998
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4011..4279
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4306..4905
                     /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    4912..5500
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5574..5807
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5879..6000
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6019..6154
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6192..6208)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6216..6232
                     /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(6240..6270)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6285..6306
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6594..7182)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7353..8213)
                     /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(8214..8318)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8792..8808
                     /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 atggcttcag cagcacgctt gacaatgatg tgggaggagg
     2341 tcacatgccc tatctgcctg gaccccttcg tggagcctgt gagcatcgag tgtggccaca
     2401 gcttctgcca ggaatgcatc tctcaggttg ggaaaggtgg gggcagcgtc tgtcctgtgt
     2461 gccggcagcg ctttctgctc aagaatctcc ggcccaatcg acagctagcc aacatggtga
     2521 acaaccttaa agaaatcagc caggaggcca gagagggcac acagggggaa cggtgtgcag
     2581 tgcatggaga gagacttcac ctgttctgtg agaaagatgg gaaggccctt tgctgggtat
     2641 gtgcccagtc tcggaaacac cgtgaccacg ccatggtccc tcttgaggag gctgcacagg
     2701 agtaccagga gaagctccag gtggcattag gggaactgag aagaaagcag gagttggctg
     2761 agaagttgga agtggaaatt gcaataaaga gagcagactg gaagaaaaca gtggaaacac
     2821 agaaatctag gattcacgca gagtttgtgc agcaaaaaaa cttcctggtt gaagaagaac
     2881 agaggcagct gcaggagctg gagaaggatg agagggagca gctgagaatc ctgggggaga
     2941 aagaggccaa gctggcccag cagagccagg ccctacagga gctcatctca gagctagatc
     3001 gaaggtgcca cagctcagca ctggaactgc tgcaggaggt gataattgtc ctggaaagga
     3061 gtgagtcctg gaacctgaag gacctggata ttacctctcc agaactcagg agtgtgtgcc
     3121 atgtgccagg gctgaagaag atgctgagga catgtgcagt ccacatcact ctggatccag
     3181 acacagccaa tccgtggctg atactttcag aagatcggag acaagtgagg cttggagaca
     3241 cccagcagag catacctgga aatgaagaga gatttgatag ttatcctatg gtcctgggtg
     3301 cccagcactt tcactctgga aaacattact gggaggtaga tgtgacagga aaggaggcct
     3361 gggacctggg tgtctgcaga gactctgtgc gcaggaaggg gcactttttg cttagttcca
     3421 agagtggctt ctggacaatt tggttgtgga acaaacaaaa atatgaggct ggcacctacc
     3481 cccagactcc cctccacctt caggtgcctc catgccaagt tgggattttc ctggactatg
     3541 aggctggcat ggtctccttc tacaacatca ctgaccatgg ctccctcatc tactccttct
     3601 ctgaatgtgc ctttacagga cctctgcggc ccttcttcag tcctggtttc aatgatggag
     3661 gaaaaaacac agcccctcta accctctgtc cactgaatat tggatcacaa ggatccactg
     3721 actatcacca tcaccatcac cattagcggc cgcgaaggat ctgcgatcgc tccggtgccc
     3781 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3841 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3901 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3961 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     4021 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     4081 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     4141 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     4201 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     4261 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag
     4321 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     4381 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     4441 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4501 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4561 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4621 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4681 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4741 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4801 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4861 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct
     4921 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4981 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5041 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5101 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5161 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5221 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5281 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     5341 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5401 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5461 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     5521 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5581 gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt
     5641 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5701 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5761 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5821 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6001 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     6061 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6121 ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat
     6181 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6241 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     6301 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6361 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6421 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6481 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6541 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6601 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6661 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6721 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6781 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6841 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6901 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6961 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7021 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7081 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7141 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7201 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7261 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7321 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7381 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7441 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7501 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7561 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7621 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7681 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7741 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7801 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7861 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7921 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7981 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8041 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8101 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8161 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8281 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8341 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8401 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     8461 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     8521 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     8581 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     8641 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8701 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     8761 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
