LOCUS       V032367                10174 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032367
VERSION     V032367
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..4870
                     /label="ALOX15B(NM_001141)"
                     /note="ALOX15B(NM_001141)"
                     /gene="ALOX15B"
     CDS             4877..4942
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4962..5461
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5482..6081
                     /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    6095..6683
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6890..7523
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7652..7668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7676..7692
                     /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(7700..7730)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7745..7766
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8054..8642)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8813..9673)
                     /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(9674..9778)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9826..9960
                     /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 ccatggccga gttcagggtc agggtgtcca ccggagaagc
     2881 cttcggggct ggcacatggg acaaagtgtc tgtcagcatc gtggggaccc ggggagagag
     2941 ccccccactg cccctggaca atctcggcaa ggagttcact gcgggcgctg aggaggactt
     3001 ccaggtgacg ctcccggagg acgtaggccg agtgctgctg ctgcgcgtgc acaaggcgcc
     3061 cccagtgctg cccctgctgg ggcccctggc cccggatgcc tggttctgcc gctggttcca
     3121 gctgacaccg ccgcggggcg gccacctcct cttcccctgc taccagtggc tggagggggc
     3181 ggggaccctg gtgctgcagg agggtacagc caaggtgtcc tgggcagacc accaccctgt
     3241 gctccagcaa cagcgccagg aggagcttca ggcccggcag gagatgtacc agtggaaggc
     3301 ttacaaccca ggttggcctc actgcctgga tgaaaagaca gtggaagact tggagctcaa
     3361 tatcaaatac tccacagcca agaatgccaa cttttatcta caggctggct ctgcttttgc
     3421 agagatgaaa atcaaggggt tgctggaccg caaggggctc tggaggagtc tgaatgagat
     3481 gaaaaggatc ttcaacttcc ggaggacccc agcagctgag cacgcatttg agcactggca
     3541 ggaggacgcc ttcttcgcct cccagttcct gaatggtctc aaccctgtcc tgatccgccg
     3601 ctgtcactac ctcccaaaga acttccccgt cactgatgcc atggtggcct cagtgttggg
     3661 tcctgggacc agcttgcagg ctgagctaga gaagggctcc ctgttcttgg tggatcacgg
     3721 catcctctct ggcatccaga ccaatgtcat taatgggaag cctcagttct ctgcggcccc
     3781 aatgaccctg ctataccaga gcccaggctg cgggccgctg ctgcctctcg ccatccagct
     3841 cagccagacc cccggcccaa acagccccat cttcctgccc actgatgaca agtgggactg
     3901 gttgctggcc aagacctggg tgcgcaatgc cgagttctcc ttccatgagg ccctcacgca
     3961 cctgctgcac tcacatctgc tgcctgaggt cttcaccctg gctaccctgc gtcagctgcc
     4021 ccactgccac cctctcttca agctgctgat cccgcacacc cgatacaccc tgcacatcaa
     4081 cacactcgcc cgggagctgc ttatcgtgcc agggcaggtg gtggacaggt ccacaggcat
     4141 cggcattgaa ggcttctctg agttgataca gaggaacatg aagcagctga actattctct
     4201 cctgtgtctg cctgaggata tccggacccg aggagttgaa gacatcccag gctactacta
     4261 ccgtgatgat gggatgcaga tctggggtgc agtggaacgc tttgtctctg aaatcatcgg
     4321 tatctactac ccaagtgatg agtctgtcca agatgacaga gagctccagg cctgggtcag
     4381 agagatcttc tccaagggct tcctaaacca ggagagctca ggtataccct cctcactgga
     4441 gacccgggaa gccctggtgc agtatgtcac catggtgata ttcacctgct ccgccaagca
     4501 tgcggctgtc agtgcagggc agtttgactc ctgtgcttgg atgcccaacc tgccacccag
     4561 catgcagctg ccaccaccca cctccaaagg cctggcaaca tgcgagggct tcatagccac
     4621 cctcccacct gtcaatgcca catgtgatgt catccttgct ctctggttgc tgagcaagga
     4681 gcctggagac caaaggcccc tgggcaccta tccggatgag cacttcacag aggaggcccc
     4741 tcggcggagc atcgccacct tccagagccg cctggcccag atctcgaggg gcatccagga
     4801 gcggaaccag ggcctggtgc tgccctacac ctacctagac cctcccctca tcgagaacag
     4861 cgtctccatc ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga
     4921 ctacaaggat gacgatgaca agtagtctag ataattctac cgggtagggg aggcgctttt
     4981 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     5041 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     5101 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     5161 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     5221 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     5281 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     5341 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     5401 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     5461 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     5521 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5581 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5641 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5701 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5761 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5821 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5881 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5941 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     6001 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     6061 acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt acaaaatttg
     6121 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6181 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6241 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6301 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6361 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6421 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6481 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     6541 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6601 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6661 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     6721 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6781 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6841 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     6901 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6961 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     7021 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     7081 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     7141 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     7201 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     7261 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     7321 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7381 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7441 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7501 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     7561 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gttttaccgt
     7621 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     8041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     8101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     8161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     8221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9361 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9421 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9481 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9541 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9601 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9661 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9781 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca
     9841 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9901 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     9961 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
    10021 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
    10081 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt taaagaaatt
    10141 gtatttgtta aatatgtact acaaacttag tagt
//
