LOCUS       V036718                 9508 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036718
VERSION     V036718
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..4177
                     /label="BCKDHA(NM_000709)"
                     /note="BCKDHA(NM_000709)"
                     /gene="BCKDHA"
     CDS             4184..4273
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4296..4795
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4816..5415
                     /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    5429..6017
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6224..6857
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6986..7002)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7010..7026
                     /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(7034..7064)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7079..7100
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7388..7976)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8147..9007)
                     /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(9008..9112)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9160..9294
                     /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 ccatggcggt agcgatcgct gcagcgaggg tctggcggct
     2881 aaaccgtggt ttgagccagg ctgccctcct gctgctgcgg cagcctgggg ctcggggact
     2941 ggctagatct caccccccca ggcagcagca gcagttttca tctctggatg acaagcccca
     3001 gttcccaggg gcctcggcgg agtttataga taagttggaa ttcatccagc ccaacgtcat
     3061 ctctggaatc cccatctacc gcgtcatgga ccggcaaggc cagatcatca accccagcga
     3121 ggacccccac ctgccgaagg agaaggtgct gaagctctac aagagcatga cactgcttaa
     3181 caccatggac cgcatcctct atgagtctca gcggcagggc cggatctcct tctacatgac
     3241 caactatggt gaggagggca cgcacgtggg gagtgccgcc gccctggaca acacggacct
     3301 ggtgtttggc cagtaccggg aggcaggtgt gctgatgtat cgggactacc ccctggaact
     3361 attcatggcc cagtgctatg gcaacatcag tgacttgggc aaggggcgcc agatgcctgt
     3421 ccactacggc tgcaaggaac gccacttcgt cactatctcc tctccactgg ccacgcagat
     3481 ccctcaggcg gtgggggcgg cgtacgcagc caagcgggcc aatgccaaca gggtcgtcat
     3541 ctgttacttc ggcgaggggg cagccagtga gggggacgcc catgccggct tcaacttcgc
     3601 tgccacactt gagtgcccca tcatcttctt ctgccggaac aatggctacg ccatctccac
     3661 gcccacctct gagcagtatc gcggcgatgg cattgcagca cgaggccccg ggtatggcat
     3721 catgtcaatc cgcgtggatg gtaatgatgt gtttgccgta tacaacgcca caaaggaggc
     3781 ccgacggcgg gctgtggcag agaaccagcc cttcctcatc gaggccatga cctacaggat
     3841 cgggcaccac agcaccagtg acgacagttc agcgtaccgc tcggtggatg aggtcaatta
     3901 ctgggataaa caggaccacc ccatctcccg gctgcggcac tatctgctga gccaaggctg
     3961 gtgggatgag gagcaggaga aggcctggag gaagcagtcc cgcaggaagg tgatggaggc
     4021 ctttgagcag gccgagcgga agcccaaacc caaccccaac ctactcttct cagacgtgta
     4081 tcaggagatg cccgcccagc tccgcaagca gcaggagtct ctggcccgcc acctgcagac
     4141 ctacggggag cactacccac tggatcactt cgataaggga tccgaacaaa aactaatatc
     4201 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc
     4261 cgaggaggac ttatagtgat ctagataatt ctaccgggta ggggaggcgc ttttcccaag
     4321 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     4381 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     4441 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     4501 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     4561 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     4621 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     4681 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     4741 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     4801 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4861 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4921 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4981 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5041 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5101 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5161 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5221 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5281 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5341 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5401 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag
     5461 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5521 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5581 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5641 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5701 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5761 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5821 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5881 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5941 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6001 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca
     6061 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     6121 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     6181 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac
     6241 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     6301 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     6361 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     6421 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     6481 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     6541 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     6601 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     6661 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6721 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6781 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6841 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6901 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct
     6961 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7021 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7081 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9121 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     9181 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9241 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     9301 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     9361 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     9421 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt
     9481 gttaaatatg tactacaaac ttagtagt
//
