LOCUS       V026533                 8760 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026533
VERSION     V026533
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    1..118
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        170..473
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        474..677
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             775..801
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             814..1842
                     /label="HOXD13(NM_000523)"
                     /note="HOXD13(NM_000523)"
                     /gene="HOXD13"
     CDS             1867..1938
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        1956..2455
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             2476..3075
                     /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    3089..3677
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             3752..3985
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        4190..4294
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4295..5155
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      5326..5914
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6325..6346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6361..6391
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6399..6415
                     /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"
     primer_bind     6423..6439
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     LTR             6700..7333
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    7380..7505
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    8002..8235
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8420..8464
                     /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"
ORIGIN
        1 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
       61 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      121 ggtttattac agggacagca gagatccagt ttggactagt ggagttccgc gttacataac
      181 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      421 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      481 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      541 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      601 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      661 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
      721 gttttgacct ccatagaaga caccggcggc cacgcgtgcg gccaagccac catgtaccca
      781 tacgatgttc cagattacgc tggatccgct agcatgagcc gcgccgggag ctgggacatg
      841 gacgggctgc gggcagacgg cgggggcgcc ggtggcgccc cggcctcttc ctcctcctca
      901 tcggtggcgg cggcggcggc gtcaggccag tgccgcggct ttctctccgc gcctgtgttc
      961 gccgggacgc attcggggcg ggcggcggcg gcggcagcgg cggctgcggc ggcggcggcg
     1021 gcagcctccg gctttgcgta ccccgggacc tctgagcgca cgggctcttc ctcgtcgtcg
     1081 tcctcttctg ccgttgtagc ggcgcgcccg gaggctcccc cagccaaaga gtgcccagca
     1141 cccacgcctg cagcggccgc tgcagcgccc ccgagcgctc cagcgctggg ctacggctac
     1201 cacttcggca acggctacta cagctgccgt atgtcgcacg gcgtgggctt acagcagaat
     1261 gcgctcaagt catcgccgca cgcctcgctg ggaggctttc ccgtggagaa gtacatggac
     1321 gtgtcaggcc tggcgagcag cagcgtaccg gccaacgagg tgccagcgcg agccaaggag
     1381 gtatccttct accagggcta tacgagccct taccagcacg tgcccggcta tatcgacatg
     1441 gtgtccactt tcggctccgg ggagcctcgg cacgaggcct acatctccat ggaggggtac
     1501 cagtcctgga cgctggctaa cgggtggaac agccaggtgt actgcaccaa ggaccagcca
     1561 caggggtccc acttttggaa atcttccttt ccaggggatg tggctctaaa tcagccggac
     1621 atgtgcgtct accgaagagg gaggaagaag agagtgcctt acaccaaact gcagcttaaa
     1681 gaactggaga acgagtatgc cattaacaaa ttcattaaca aggacaagcg gcggcgtatc
     1741 tcggctgcta cgaacctatc tgagagacaa gtgaccattt ggtttcagaa ccgaagagtg
     1801 aaggacaaga aaattgtctc caagctcaaa gatactgtct ccgactcgag aagcggccgc
     1861 gtcgaggatt acaaggatga cgacgataag gactataagg acgatgatga caaggactac
     1921 aaagatgatg acgataaata gtcgagaatt ctaccgggta ggggaggcgc ttttcccaag
     1981 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     2041 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     2101 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     2161 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     2221 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     2281 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     2341 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     2401 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     2461 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     2521 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     2581 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     2641 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     2701 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     2761 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     2821 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     2881 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     2941 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3001 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3061 aagcccggtg cctgaatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag
     3121 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3181 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3241 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3301 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3361 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3421 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3481 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3541 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3601 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3661 ttgggccgcc tccccgcctg agatccttta agaccaatga cttacaaggc agctgtagat
     3721 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     3781 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     3841 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3901 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3961 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     4021 taacttgcaa agaaatgaat atcagagagt gagaggcccg ggttaattaa ggaaagggct
     4081 agatcattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4141 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4201 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4261 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4321 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4381 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4441 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4501 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4561 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4621 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4681 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4741 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4801 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4861 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4921 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4981 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5041 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5101 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5161 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5221 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5281 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5341 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5401 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5461 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5521 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5581 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5641 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5701 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5761 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5821 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5881 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5941 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6001 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6061 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6121 ccccgcgcgt tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt
     6181 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6241 tttggaggcc taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa
     6301 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6361 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6421 cacaggaaac agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca
     6481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     6541 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     6601 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     6661 aaattgtatt tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc
     6721 aaagaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     6781 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     6841 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     6901 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     6961 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     7021 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     7081 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     7141 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7201 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7261 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7321 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     7381 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     7441 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     7501 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     7561 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     7621 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     7681 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     7741 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     7801 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt
     7861 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     7921 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     7981 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     8041 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     8101 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     8161 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     8221 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     8281 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     8341 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     8401 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     8461 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     8521 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     8581 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     8641 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     8701 agtgaacgga tctcgacggt atcgccgaat taattcacaa atggcagtat tcatccacaa
//
