LOCUS       V039578                 9568 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039578
VERSION     V039578
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             2843..3559
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3629..4261
                     /label="BAG2(NM_004282)"
                     /note="BAG2(NM_004282)"
                     /gene="BAG2"
     CDS             4271..4336
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4356..4855
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4876..5475
                     /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    5489..6077
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6284..6917
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7046..7062)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7070..7086
                     /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(7094..7124)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7139..7160
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7448..8036)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8207..9067)
                     /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(9068..9172)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9220..9354
                     /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 ctcaagcttc gaattcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg
     3601 gggaagcgga ggcggaggga gtggatccat ggctcaggcg aagatcaacg ctaaagccaa
     3661 cgaggggcgc ttctgccgct cctcctccat ggctgaccgc tccagccgcc tgctggagag
     3721 cctggaccag ctggagctca gggttgaagc tttgagagaa gcagcaactg ctgttgagca
     3781 agagaaagaa atccttctgg aaatgatcca cagtatccaa aatagccagg acatgaggca
     3841 gatcagtgac ggagaaagag aagaattaaa tctgactgca aaccgtttga tgggaagaac
     3901 tctcaccgtt gaagtgtcag tagaaacaat tagaaacccc cagcagcaag aatccctaaa
     3961 gcatgccaca aggattattg atgaggtggt caataagttt ctggatgatt tgggaaatgc
     4021 caagagtcat ttaatgtcgc tctacagtgc atgttcatct gaggtgccac atgggccagt
     4081 tgatcagaag tttcaatcca tagtaattgg ctgtgctctt gaagatcaga agaaaattaa
     4141 gagaagatta gagactctgc ttagaaatat tgaaaactct gacaaggcca tcaagctatt
     4201 agagcattct aaaggagctg gttccaaaac tctgcaacaa aatgctgaaa gcagattcaa
     4261 tggcggccgc gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     4321 ggatgacgat gacaagtagt ctagataatt ctaccgggta ggggaggcgc ttttcccaag
     4381 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     4441 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     4501 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     4561 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     4621 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     4681 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     4741 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     4801 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     4861 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4921 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4981 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5041 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5101 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5161 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5221 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5281 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5341 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5401 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5461 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag
     5521 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5581 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5641 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5701 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5761 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5821 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5881 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5941 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6001 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6061 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca
     6121 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     6181 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     6241 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac
     6301 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     6361 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     6421 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     6481 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     6541 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     6601 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     6661 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     6721 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6781 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6841 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6901 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6961 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct
     7021 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7081 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7141 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7201 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7861 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9181 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     9241 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9301 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     9361 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     9421 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     9481 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt
     9541 gttaaatatg tactacaaac ttagtagt
//
