LOCUS       V020128                 9274 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020128
VERSION     V020128
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..4004
                     /label="MAP2K1(NM_002755)"
                     /note="MAP2K1(NM_002755)"
                     /gene="MAP2K1"
     CDS             4005..4031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4062..4561
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4582..5181
                     /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    5195..5783
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5990..6623
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6752..6768)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6776..6792
                     /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(6800..6830)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6845..6866
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7154..7742)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7913..8773)
                     /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(8774..8878)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8926..9060
                     /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 ccaccatgcc caagaagaag ccgacgccca tccagctgaa cccggccccc gacggctctg
     2881 cagttaacgg gaccagctct gcggagacca acttggaggc cttgcagaag aagctggagg
     2941 agctagagct tgatgagcag cagcgaaagc gccttgaggc ctttcttacc cagaagcaga
     3001 aggtgggaga actgaaggat gacgactttg agaagatcag tgagctgggg gctggcaatg
     3061 gcggtgtggt gttcaaggtc tcccacaagc cttctggcct ggtcatggcc agaaagctaa
     3121 ttcatctgga gatcaaaccc gcaatccgga accagatcat aagggagctg caggttctgc
     3181 atgagtgcaa ctctccgtac atcgtgggct tctatggtgc gttctacagc gatggcgaga
     3241 tcagtatctg catggagcac atggatggag gttctctgga tcaagtcctg aagaaagctg
     3301 gaagaattcc tgaacaaatt ttaggaaaag ttagcattgc tgtaataaaa ggcctgacat
     3361 atctgaggga gaagcacaag atcatgcaca gagatgtcaa gccctccaac atcctagtca
     3421 actcccgtgg ggagatcaag ctctgtgact ttggggtcag cgggcagctc atcgactcca
     3481 tggccaactc cttcgtgggc acaaggtcct acatgtcgcc agaaagactc caggggactc
     3541 attactctgt gcagtcagac atctggagca tgggactgtc tctggtagag atggcggttg
     3601 ggaggtatcc catccctcct ccagatgcca aggagctgga gctgatgttt gggtgccagg
     3661 tggaaggaga tgcggctgag accccaccca ggccaaggac ccccgggagg ccccttagct
     3721 catacggaat ggacagccga cctcccatgg caatttttga gttgttggat tacatagtca
     3781 acgagcctcc tccaaaactg cccagtggag tgttcagtct ggaatttcaa gattttgtga
     3841 ataaatgctt aataaaaaac cccgcagaga gagcagattt gaagcaactc atggttcatg
     3901 cttttatcaa gagatctgat gctgaggaag tggattttgc aggttggctc tgctccacca
     3961 tcggccttaa ccagcccagc acaccaaccc atgctgctgg cgtctaccca tacgacgtac
     4021 cagattacgc ttaggatccc gcgactctag ataattctac cgggtagggg aggcgctttt
     4081 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     4141 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     4201 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     4261 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     4321 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     4381 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     4441 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     4501 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     4561 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5161 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg
     5221 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5281 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5341 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5401 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5461 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5521 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5581 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5641 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5701 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5761 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     5821 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5881 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5941 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     6001 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6061 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     6121 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     6181 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     6241 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     6301 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     6361 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     6421 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6481 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6541 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6601 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6661 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt
     6721 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6781 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6841 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6901 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8881 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca
     8941 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9001 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     9061 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
     9121 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
     9181 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt
     9241 gtatttgtta aatatgtact acaaacttag tagt
//
