LOCUS       V020921                 9518 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020921
VERSION     V020921
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..4214
                     /label="CTSC(NM_001814)"
                     /note="CTSC(NM_001814)"
                     /gene="CTSC"
     CDS             4221..4286
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4306..4805
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4826..5425
                     /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    5439..6027
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6234..6867
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6996..7012)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7020..7036
                     /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(7044..7074)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7089..7110
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7398..7986)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8157..9017)
                     /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(9018..9122)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9170..9304
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatggg tgctgggccc tccttgctgc tcgccgccct cctgctgctt ctctccggcg
     2881 acggcgccgt gcgctgcgac acacctgcca actgcaccta tcttgacctg ctgggcacct
     2941 gggtcttcca ggtgggctcc agcggttccc agcgcgatgt caactgctcg gttatgggac
     3001 cacaagaaaa aaaagtagtg gtgtaccttc agaagctgga tacagcatat gatgaccttg
     3061 gcaattctgg ccatttcacc atcatttaca accaaggctt tgagattgtg ttgaatgact
     3121 acaagtggtt tgcctttttt aagtataaag aagagggcag caaggtgacc acttactgca
     3181 acgagacaat gactgggtgg gtgcatgatg tgttgggccg gaactgggct tgtttcaccg
     3241 gaaagaaggt gggaactgcc tctgagaatg tgtatgtcaa catagcacac cttaagaatt
     3301 ctcaggaaaa gtattctaat aggctctaca agtatgatca caactttgtg aaagctatca
     3361 atgccattca gaagtcttgg actgcaacta catacatgga atatgagact cttaccctgg
     3421 gagatatgat taggagaagt ggtggccaca gtcgaaaaat cccaaggccc aaacctgcac
     3481 cactgactgc tgaaatacag caaaagattt tgcatttgcc aacatcttgg gactggagaa
     3541 atgttcatgg tatcaatttt gtcagtcctg ttcgaaacca agcatcctgt ggcagctgct
     3601 actcatttgc ttctatgggt atgctagaag cgagaatccg tatactaacc aacaattctc
     3661 agaccccaat cctaagccct caggaggttg tgtcttgtag ccagtatgct caaggctgtg
     3721 aaggcggctt cccatacctt attgcaggaa agtacgccca agattttggg ctggtggaag
     3781 aagcttgctt cccctacaca ggcactgatt ctccatgcaa aatgaaggaa gactgctttc
     3841 gttattactc ctctgagtac cactatgtag gaggtttcta tggaggctgc aatgaagccc
     3901 tgatgaagct tgagttggtc catcatgggc ccatggcagt tgcttttgaa gtatatgatg
     3961 acttcctcca ctacaaaaag gggatctacc accacactgg tctaagagac cctttcaacc
     4021 cctttgagct gactaatcat gctgttctgc ttgtgggcta tggcactgac tcagcctctg
     4081 ggatggatta ctggattgtt aaaaacagct ggggcaccgg ctggggtgag aatggctact
     4141 tccggatccg cagaggaact gatgagtgtg caattgagag catagcagtg gcagccacac
     4201 caattcctaa attgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     4261 tcgactacaa ggatgacgat gacaagtaat ctagataatt ctaccgggta ggggaggcgc
     4321 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     4381 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     4441 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     4501 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     4561 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     4621 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     4681 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     4741 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     4801 tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     4861 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4921 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4981 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5041 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5101 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5161 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5221 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5281 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5341 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5401 catgacccgc aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa
     5461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5821 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     5881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     5941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6001 ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa
     6061 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     6121 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     6181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg
     6241 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     6301 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
     6361 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
     6421 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     6481 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
     6541 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
     6601 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
     6661 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6721 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6781 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6841 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6901 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta
     6961 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7021 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7081 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7141 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7201 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7801 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     8221 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     8281 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     8341 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8401 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8461 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8521 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8581 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8641 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8701 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8761 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8821 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8881 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8941 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9001 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9061 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9121 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
     9181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     9241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     9301 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
     9361 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
     9421 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga
     9481 aattgtattt gttaaatatg tactacaaac ttagtagt
//
