LOCUS       V021884                 9978 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021884
VERSION     V021884
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             2827..4704
                     /label="NXF2(NM_022053)"
                     /note="NXF2(NM_022053)"
                     /gene="NXF2"
     CDS             4711..4734
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4766..5265
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5286..5885
                     /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    5899..6487
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6694..7327
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7456..7472)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7480..7496
                     /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(7504..7534)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7549..7570
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7858..8446)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8617..9477)
                     /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(9478..9582)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9630..9764
                     /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 gccaccatgt gctctactct aaagaagtgt gggacataca gaactgaagt tgcagaatgc
     2881 catgaccatg gtagcacttt tcaaggaaga aagaaaggtg ggagttcttt ccgggataat
     2941 tttgacaaga ggagctgtca ttatgaacat ggtgggtatg agcgcccgcc ttcacactgc
     3001 caggagaatg atggaagcgt ggagatgagg gatgtccaca aggaccaaca actaagacac
     3061 actccttata gcatccgatg cgaaagaaga atgaaatggc atagtgaaga cgaaatccgt
     3121 attaccacgt ggagaaatag aaaacctccg gagagaaaaa tgagtcagaa cacacaggat
     3181 ggatacacaa ggaactggtt taaggtcaca attccttacg ggataaagta tgacaaggca
     3241 tggctaatga attcaatcca gagccattgc agtgaccgct tcactccggt tgatttccac
     3301 tacgtccgaa atcgggcatg cttctttgtc caggatgcta gcgctgcctc cgcattgaag
     3361 gatgtcagtt ataagattta tgatgatgag aaccaaaaga tatgtatatt tgtcaatcat
     3421 tctactgcgc cctactctgt gaagaataag ttgaagccag gccaaatgga gatgctaaag
     3481 ctgaccatga acaaacggta caatgtctcc cagcaagctc ttgatctcca gaatctccgc
     3541 tttgacccag acttgatggg ccgtgacatt gatataatcc tgaatcgaag aaactgcatg
     3601 gctgccaccc tgaagatcat tgaaagaaat ttccctgagc tgttgtcttt gaacttgtgc
     3661 aacaacaagc tgtaccagct ggatggcctt tctgacatta cagagaaggc tcccaaagtc
     3721 aagaccctga atctctccaa aaataagctg gagtcggcgt gggagttggg caaggtgaaa
     3781 gggctgaagc tcgaagagct atggctagaa gggaacccgt tgtgcagcac cttctcggac
     3841 cagtccgcct atgtaagtgc catccgggat tgtttcccca agttgttacg cctggacggc
     3901 cgagagttat ccgcaccagt gattgttgac attgacagct ctgagacaat gaaaccctgc
     3961 aaggaaaact ttactggatc tgagacccta aagcatttag tcctgcaatt cctgcagcag
     4021 tattactcga tctatgactc tggagatcga cagggtctcc tcggtgctta ccacgatgag
     4081 gcctgcttct ccttggctat tcccttcgac cccaaggact cagccccgag cagcttgtgc
     4141 aagtactttg aggatagcag gaatatgaaa acactcaagg acccctacct gaagggggaa
     4201 ctgctgaggc gcacaaaacg tgacattgtg gactccctca gtgcgttgcc caaaactcag
     4261 catgacctca gctccatcct ggtggacgtg tggtgccaga cggaaaggat gctctgcttt
     4321 tctgtcaatg gggttttcaa ggaagtggaa ggacagtctc agggttctgt tctcgccttc
     4381 acccggacct tcattgctac ccctggcagc agttccagtc tgtgcatcgt gaatgacgag
     4441 ctgtttgtga gggatgccag cccccaagag actcagagtg ccttctccat cccagtgtcc
     4501 acactctcct ccagctctga gccctccctc tcccaggagc agcaggaaat ggtgcaggct
     4561 ttctctgccc agtctgggat gaaactggag tggtctcaga agtgccttca ggacaatgag
     4621 tggaactaca ctagagctgg ccaggccttc actatgctcc agaccgaggg caagatcccc
     4681 gcggaggcct tcaagcaaat ctccgaattc gattacaagg atgacgacga taagtgagga
     4741 tcccgcgact ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     4801 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     4861 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     4921 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     4981 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     5041 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     5101 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     5161 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     5221 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc
     5281 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5341 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5401 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5461 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5521 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5581 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5641 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5701 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5761 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5821 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5881 cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5941 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6001 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6061 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6121 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6181 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     6301 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6361 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6421 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6481 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
     6541 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     6601 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     6661 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
     6721 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     6781 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     6841 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     6901 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     6961 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     7021 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     7081 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     7141 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7201 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7261 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7321 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7381 atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag
     7441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9601 gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca
     9661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     9721 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag
     9781 ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt
     9841 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
     9901 tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg
     9961 tactacaaac ttagtagt
//
