LOCUS       V039219                 9358 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039219
VERSION     V039219
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..4060
                     /label="BHMT(NM_001713)"
                     /note="BHMT(NM_001713)"
                     /gene="BHMT"
     CDS             4061..4126
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4146..4645
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4666..5265
                     /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    5279..5867
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6074..6707
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6836..6852)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6860..6876
                     /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(6884..6914)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6929..6950
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7238..7826)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7997..8857)
                     /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(8858..8962)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9010..9144
                     /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 ccatgccacc cgttgggggc aaaaaggcca agaagggcat
     2881 cctagaacgt ttaaatgctg gagagattgt gattggagat ggagggtttg tctttgcact
     2941 ggagaagagg ggctacgtaa aggcaggacc ctggactcct gaagctgctg tggagcaccc
     3001 agaagcagtt cgccagcttc atcgagagtt cctcagagct ggctcaaacg tcatgcagac
     3061 cttcaccttc tatgcgagtg aagacaagct ggagaacagg ggcaactatg tcttagagaa
     3121 gatatctggg caggaagtca atgaagctgc ttgcgacatc gcccgacaag tggctgatga
     3181 aggagatgct ttggtagcag gaggagtgag tcagacacct tcatacctta gctgcaagag
     3241 tgaaactgaa gtcaaaaaag tatttctgca acagttagag gtctttatga agaagaacgt
     3301 ggacttcttg attgcagagt attttgaaca cgttgaagaa gctgtgtggg cagttgaaac
     3361 cttgatagca tccggtaaac ctgtggcagc aaccatgtgc attggcccag aaggagattt
     3421 gcatggcgtg ccccccggcg agtgtgcagt gcgcctggtg aaagcaggag catccatcat
     3481 tggtgtgaac tgccactttg accccaccat tagtttaaaa acagtgaagc tcatgaagga
     3541 gggcttggag gctgcccgac tgaaagctca cctgatgagc cagcccttgg cttaccacac
     3601 tcctgactgc aacaagcagg gattcatcga tctcccagaa ttcccatttg gactggaacc
     3661 cagagttgcc accagatggg atattcaaaa atacgccaga gaggcctaca acctgggggt
     3721 caggtacatt ggcgggtgct gtggatttga gccctaccac atcagggcaa ttgcagagga
     3781 gctggcccca gaaaggggct ttttgccacc agcttcagaa aaacatggca gctggggaag
     3841 tggtttggac atgcacacca aaccctgggt tagagcaagg gccaggaagg aatactggga
     3901 gaatcttcgg atagcctcag gccggccata caacccttca atgtcaaagc cagatggctg
     3961 gggagtgacc aaaggaacag ccgagctgat gcagcagaaa gaagccacaa ctgagcagca
     4021 gctgaaagag ctctttgaaa aacaaaaatt caaatcacag gactacaaag accatgacgg
     4081 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt ctagataatt
     4141 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     4201 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     4261 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     4321 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     4381 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     4441 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     4501 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     4561 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     4621 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
     4681 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     4741 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4801 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4861 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4921 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4981 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     5041 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5101 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5161 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5221 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaccgcg tctggaacaa
     5281 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5341 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5401 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5461 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5521 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5581 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5641 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     5701 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5761 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5821 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc
     5881 tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     5941 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6001 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6061 gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6121 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg
     6181 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag
     6241 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg
     6301 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat
     6361 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac
     6421 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
     6481 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt
     6541 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6601 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6661 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     6721 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc
     6781 ttgacattgc tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6841 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6901 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6961 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7021 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7081 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7141 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     9001 cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     9061 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     9121 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     9181 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     9241 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     9301 aacttagtag tttttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
