LOCUS       V039206                 9666 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039206
VERSION     V039206
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    2058..2175
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2227..2530
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2531..2734
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2854..4815
                     /label="HSPA5(NM_005347)"
                     /note="HSPA5(NM_005347)"
                     /gene="HSPA5"
     CDS             4834..4905
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        4923..5422
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5443..6042
                     /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    6056..6644
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6719..6952
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        7157..7261
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7262..8122
                     /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      8293..8881
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    9292..9313
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        9328..9358
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    9366..9382
                     /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     9390..9406
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
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 tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccac gcgtcgaagc
     2821 ccgggcggat cgggtttaaa cggatccgcc accatgaagc tctccctggt ggccgcgatg
     2881 ctgctgctgc tcagcgcggc gcgggccgag gaggaggaca agaaggagga cgtgggcacg
     2941 gtggtcggca tcgacctggg gaccacctac tcctgcgtcg gcgtgttcaa gaacggccgc
     3001 gtggagatca tcgccaacga tcagggcaac cgcatcacgc cgtcctatgt cgccttcact
     3061 cctgaagggg aacgtctgat tggcgatgcc gccaagaacc agctcacctc caaccccgag
     3121 aacacggtct ttgacgccaa gcggctcatc ggccgcacgt ggaatgaccc gtctgtgcag
     3181 caggacatca agttcttgcc gttcaaggtg gttgaaaaga aaactaaacc atacattcaa
     3241 gttgatattg gaggtgggca aacaaagaca tttgctcctg aagaaatttc tgccatggtt
     3301 ctcactaaaa tgaaagaaac cgctgaggct tatttgggaa agaaggttac ccatgcagtt
     3361 gttactgtac cagcctattt taatgatgcc caacgccaag caaccaaaga cgctggaact
     3421 attgctggcc taaatgttat gaggatcatc aacgagccta cggcagctgc tattgcttat
     3481 ggcctggata agagggaggg ggagaagaac atcctggtgt ttgacctggg tggcggaacc
     3541 ttcgatgtgt ctcttctcac cattgacaat ggtgtcttcg aagttgtggc cactaatgga
     3601 gatactcatc tgggtggaga agactttgac cagcgtgtca tggaacactt catcaaactg
     3661 tacaaaaaga agacgggcaa agatgtcagg aaagacaata gagctgtgca gaaactccgg
     3721 cgcgaggtag aaaaggccaa acgggccctg tcttctcagc atcaagcaag aattgaaatt
     3781 gagtccttct atgaaggaga agacttttct gagaccctga ctcgggccaa atttgaagag
     3841 ctcaacatgg atctgttccg gtctactatg aagcccgtcc agaaagtgtt ggaagattct
     3901 gatttgaaga agtctgatat tgatgaaatt gttcttgttg gtggctcgac tcgaattcca
     3961 aagattcagc aactggttaa agagttcttc aatggcaagg aaccatcccg tggcataaac
     4021 ccagatgaag ctgtagcgta tggtgctgct gtccaggctg gtgtgctctc tggtgatcaa
     4081 gatacaggtg acctggtact gcttgatgta tgtcccctta cacttggtat tgaaactgtg
     4141 ggaggtgtca tgaccaaact gattccaagg aacacagtgg tgcctaccaa gaagtctcag
     4201 atcttttcta cagcttctga taatcaacca actgttacaa tcaaggtcta tgaaggtgaa
     4261 agacccctga caaaagacaa tcatcttctg ggtacatttg atctgactgg aattcctcct
     4321 gctcctcgtg gggtcccaca gattgaagtc acctttgaga tagatgtgaa tggtattctt
     4381 cgagtgacag ctgaagacaa gggtacaggg aacaaaaata agatcacaat caccaatgac
     4441 cagaatcgcc tgacacctga agaaatcgaa aggatggtta atgatgctga gaagtttgct
     4501 gaggaagaca aaaagctcaa ggagcgcatt gatactagaa atgagttgga aagctatgcc
     4561 tattctctaa agaatcagat tggagataaa gaaaagctgg gaggtaaact ttcctctgaa
     4621 gataaggaga ccatggaaaa agctgtagaa gaaaagattg aatggctgga aagccaccaa
     4681 gatgctgaca ttgaagactt caaagctaag aagaaggaac tggaagaaat tgttcaacca
     4741 attatcagca aactctatgg aagtgcaggc cctcccccaa ctggtgaaga ggatacagca
     4801 gaaaaagatg agttgctcga gtctagaggg cccgattaca aggatgacga cgataaggat
     4861 tacaaggatg acgacgataa ggattacaag gatgacgacg ataagtagtc gagaattcta
     4921 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     4981 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     5041 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     5101 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     5161 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     5221 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     5281 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     5341 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     5401 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     5461 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5521 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5581 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5641 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5701 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5761 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5821 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5881 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5941 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     6001 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaatcgatag atcctaatca
     6061 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6121 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6181 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6241 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6301 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6361 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6421 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6481 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6541 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6601 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgaga tcctttaaga
     6661 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     6721 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     6781 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     6841 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     6901 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     6961 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     7021 aggcccgggt taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat
     7081 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     7141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7201 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7261 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7321 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7381 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7441 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7501 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7561 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7621 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7681 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7741 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7801 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7861 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7921 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7981 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8041 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8101 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8161 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8221 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8401 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8941 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     9001 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     9061 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa
     9121 gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     9181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc
     9241 gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     9301 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     9361 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt
     9421 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9481 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     9541 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     9601 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     9661 agtagt
//
