LOCUS       V043563                 9125 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043563
VERSION     V043563
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"
     regulatory      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..4027
                     /label="Hsf1(NM_001331152)"
                     /note="Hsf1(NM_001331152)"
                     /gene="Hsf1"
     promoter        4095..4306
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4319..4587
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4614..5213
                     /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    5220..5808
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5882..6115
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6187..6308
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6327..6462
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6500..6516)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6524..6540
                     /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(6548..6578)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6593..6614
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6902..7490)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7661..8521)
                     /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(8522..8626)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9100..9116
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat ggatctggcc gtgggccccg gtgcagcggg
     2401 gcccagcaac gtcccggcct tcctaaccaa gctgtggacc ctcgtgagcg acccggacac
     2461 agacgcgctc atctgctgga gcccgagtgg gaacagcttc cacgtgtttg accagggcca
     2521 gtttgccaag gaggtgctgc ccaagtactt caagcacaac aacatggcta gcttcgtgcg
     2581 gcagctcaac atgtatggct tccgaaaagt agtccacatt gagcagggtg gcctggtcaa
     2641 gcctgagaga gatgacaccg agttccagca tccttgtttc ttgcgtggac aggaacagct
     2701 ccttgagaac atcaagagga aagtgaccag cgtgtccacc ctgaagagtg aggacataaa
     2761 aatacgccag gacagtgtca cccggctgtt gacagatgtg cagctgatga aggggaaaca
     2821 ggagtgtatg gactccaagc tcctggccat gaagcacgag aacgaggccc tgtggcggga
     2881 ggtggccagc cttcggcaga agcatgccca gcagcaaaaa gttgtcaaca agctcattca
     2941 gttcctgatc tcactggtgc agtcgaaccg gatcctgggg gtgaagagaa agatccctct
     3001 gatgttgagt gacagcaact cagcacactc tgtgcccaag tatggtcgac agtactccct
     3061 ggagcatgtc catggtcctg gcccatactc agctccatct ccagcctaca gcagctctag
     3121 cctttactcc tctgatgctg tcaccagctc tggacccata atctccgata tcactgagct
     3181 ggctcccacc agccctttgg cctccccagg caggagcata gatgagaggc ctctgtccag
     3241 cagcactctg gtccgtgtca agcaagagcc ccccagccca cctcacagcc ctcgggtact
     3301 ggaggcgagc cctgggcgcc catcctccat ggatacccct ttgtccccaa ctgccttcat
     3361 tgactccatc cttcgagaga gcgagcctac ccctgctgcc tcaaacacag cccctatgga
     3421 cacaaccgga gcccaagccc ccgcactccc gaccccctcc acccctgaga agtgcctcag
     3481 cgtagcctgc ctagacaatt tggctcgcgc tccacagatg tctggggtcg cccgcctctt
     3541 cccctgcccc tcttcctttc tgcatggccg agtccagcca gggaacgagc taagtgatca
     3601 cctggatgcc atggactcca acctggacaa cctgcagacc atgctgacaa gccacggctt
     3661 cagtgtggac accagtgccc tgctggacct gttcagcccc tcggtgacca tgcccgacat
     3721 gagcctgcct gacctggaca gcagcctggc cagcattcag gagcttctgt ctccacaaga
     3781 gcctcccagg cctattgagg cagagaacag taaccccgac tcaggaaagc agctggtgca
     3841 ctacacggct cagcctctgt tcctgctgga tcctgatgct gtggacacag ggagcagtga
     3901 gctgcctgtg ctctttgagc tgggggagag ctcctacttc tctgaggggg atgactacac
     3961 ggatgatccc accatctctc ttctgacagg cactgaaccc cataaagcca aggaccccac
     4021 tgtctcctag ggatccctcg agagatcagt gtagcggccg cgaaggatct gcgatcgctc
     4081 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4141 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4201 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     4261 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     4321 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     4381 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     4441 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     4501 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     4561 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg
     4621 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     4681 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     4741 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4801 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4861 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4921 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4981 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5041 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5101 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5161 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca
     5221 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5281 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5341 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5401 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5461 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5521 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5581 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5641 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5701 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5761 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     5821 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5881 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     5941 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6001 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6061 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6121 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6181 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6241 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6301 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6481 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6541 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6601 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6661 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6721 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6781 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6841 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6901 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6961 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7021 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7081 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7141 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7201 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7261 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7321 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7381 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7441 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7501 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7561 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7621 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7681 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7741 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7801 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7861 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7921 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7981 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8041 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8101 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8161 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8221 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8281 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8341 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8401 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8461 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8521 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8581 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8641 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8701 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8761 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8821 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8881 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8941 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9001 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9061 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9121 agctg
//
