LOCUS       V046467                 9371 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046467
VERSION     V046467
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4203
                     /label="Clpx(NM_011802)"
                     /note="Clpx(NM_011802)"
                     /gene="Clpx"
     CDS             4210..4236
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4240..4266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4270..4296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4341..4552
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4565..4833
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4860..5459
                     /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    5466..6054
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6128..6361
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6433..6554
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6573..6708
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6746..6762)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6770..6786
                     /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(6794..6824)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6839..6860
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7148..7736)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7907..8767)
                     /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(8768..8872)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9346..9362
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgtccagt tgcggcgctt gtacctgcgg cgctgcggcc
     2341 gcccggcttc tcaccacttc gctcacctcc gcgcagagag gtatttcgtg tgggcgaatc
     2401 catgtgccag tgttaggaag actggggacg accttggacg ctcaggctct gcgcagagct
     2461 cctcttagaa ccttttcaga aacaccagca tactttgcct caaaagacgg ggcaaacaag
     2521 gatggctccg gagatggaaa taagaaatca gtgactgaag gaagcagtaa gaaatcaggc
     2581 tctgggaatt ctgggaaagg tggaaaccag cttcgctgtc ctaaatgtgg tgatttgtgt
     2641 acacacgtgg agacctttgt gtcttccacg cgttttgtta agtgtgaaaa atgtcatcat
     2701 ttttttgtgg tgctgtcgga agcagactca aagaaaagca taattaagga acccgagtcc
     2761 gctgcagagg ctgtgaagtt ggcattccag cagaagccac ctccgccccc caaaaagatt
     2821 tataactacc tcgacaagta tgttgttggc cagtcgtttg ctaagaaagt gctttcagtt
     2881 gccgtgtata atcattataa gagaatatat aataatattc cagctaatct gagacagcaa
     2941 gcagaggctg agaagcagac atcactaaca cctagagagt tagaaataag aagacgggag
     3001 gatgagtaca gatttacaaa gttgcttcag atcgctggga tcagcccaca cggcaatgcc
     3061 ttgggagcct ctatgcagca gcaggtaaac cagcaaatgc ctcaggagaa aagaggaggt
     3121 gaagtgttgg actcttctca agatgatata aaactggaaa aaagtaatat tttgctgctt
     3181 ggaccaactg ggtcaggtaa aacccttctg gcacaaactt tagctaaatg ccttgatgtt
     3241 ccttttgcta tctgtgactg tacaactttg acccaggctg gatatgtagg tgaagatatt
     3301 gaatctgtga ttgccaaact actccaagat gccaattaca atgtagagaa agcacaacaa
     3361 ggaattgtct ttctggatga agtagataag attggcagtg tgccaggcat tcatcaatta
     3421 cgggatgtag gtggagaagg cgttcagcaa ggtttattaa aacttctaga aggcacaata
     3481 gtcaatgttc cagaaaagaa ttctcgaaaa ctacgtggag agacagttca agttgataca
     3541 acaaatgtct tgtttgtggc atctggtgcc tttaatggct tagacagaat catcagcagg
     3601 aggaaaaatg aaaagtatct cggctttgga acaccatcta acctgggaaa gggcagaagg
     3661 gctgcagctg cagcggatct tgctaaccga agtggggaat ctaacactca tcaggacatt
     3721 gaggagaaag accggttact ccgccatgtg gaagccagag atcttattga gtttggcatg
     3781 ataccagagt ttgtgggacg gttgcctgtg gtggttcctt tgcacagcct agatgagaag
     3841 acacttgtac aaatactgac cgagccacgg aatgctgtta ttcctcagta tcaggccttg
     3901 ttcagcatgg ataagtgtga gctgaatgtt actgaggatg ctctgaaagc catagcgaga
     3961 ttggccctgg aaagaaaaac aggtgcacga ggccttcggt ctataatgga aaagctgtta
     4021 ctagaaccaa tgttcgaagt tcctaattct gacattgtat gtgtggaagt tgacaaagaa
     4081 gtagtagaag gcaaaaagga accaggatac attcgggcac cctcaaaaga gtcctctgaa
     4141 gaggaatatg actctggagt ggaggaggat ggatggcctc gtcaagcgga tgctgcaaac
     4201 agcggatcct acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     4261 tatgctagct atccttatga tgtcccagac tacgcatagg cggccgcgaa ggatctgcga
     4321 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     4381 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4441 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4501 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     4561 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     4621 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     4681 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     4741 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     4801 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     4861 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     4921 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     4981 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5041 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5101 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5161 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5221 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5281 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5341 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5401 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     5461 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5521 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5581 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5641 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5701 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5761 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5821 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5881 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5941 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6001 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta
     6061 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6121 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     6181 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6241 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6301 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6361 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6421 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6481 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6541 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6721 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6781 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6841 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6901 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6961 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7021 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7081 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7561 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7921 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7981 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8041 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8101 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8161 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8221 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8281 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8341 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8401 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8461 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8521 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8581 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8641 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8701 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8761 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8821 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8881 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8941 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     9001 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     9061 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     9121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     9181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     9241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     9301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     9361 gtgccaagct g
//
