LOCUS       V040094                 9656 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040094
VERSION     V040094
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..4488
                     /label="Tbk1(NM_019786)"
                     /note="Tbk1(NM_019786)"
                     /gene="Tbk1"
     CDS             4495..4521
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4525..4551
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4555..4581
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4626..4837
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4850..5118
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5145..5744
                     /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    5751..6339
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6413..6646
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6718..6839
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6858..6993
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7031..7047)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7055..7071
                     /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(7079..7109)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7124..7145
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7433..8021)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8192..9052)
                     /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(9053..9157)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9631..9647
                     /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 catgcagagc acctccaacc atctgtggct cctgtctgat
     2341 atcctaggcc agggggccac tgcaaatgtc ttccgaggaa ggcataagaa aactggtgat
     2401 ctctatgctg tcaaagtatt taataacata agcttccttc gcccagtgga tgttcaaatg
     2461 agagaatttg aagtgttaaa aaaactcaat cacaaaaaca ttgtcaagtt atttgctatt
     2521 gaagaggaga caacaacaag acataaagtg cttattatgg agttttgtcc ctgtgggagt
     2581 ttatacactg ttctagagga gccgtccaat gcgtatggac ttccagaatc agaatttctc
     2641 attgtcttac gagatgtggt gggcgggatg aatcatctcc gagagaacgg catagtgcac
     2701 cgagatatca agccaggcaa catcatgcgc gtcatagggg aggacggcca gtctgtgtac
     2761 aaactcacgg atttcggcgc cgctcgagag ctggaggacg atgagcagtt tgtgtctctg
     2821 tacggcacag aagagtacct gcatccggac atgtatgaaa gggcagtgct aagaaaggac
     2881 catcagaaga agtacggggc taccgttgat ctgtggagtg ttggagtgac attctaccat
     2941 gcagccacgg ggtcgctgcc gtttagaccc ttcgaggggc ctcggaggaa caaagaagta
     3001 atgtataaaa taatcactgg gaagccgtct ggtgcaatat ctggagtaca gaaagcagaa
     3061 aacggaccaa ttgactggag tggagacatg cctctctcct gtagtctttc tcagggtctt
     3121 caggcactgc ttaccccagt tcttgcaaac atacttgaag ctgatcagga gaagtgctgg
     3181 ggttttgacc agttctttgc agagaccagt gatgtgcttc accgaatggt gatccatgtc
     3241 ttctcgctac aacacatgac ggcgcataag atttacattc acagctataa cactgctgct
     3301 gtgttccatg aactggtcta taaacaaacc aagattgttt cctcaaatca agaacttatc
     3361 tacgaaggac gacgcttagt cctagaactc ggacgactag cccagcattt tcctaaaacc
     3421 acagaggaaa atcctatctt tgtcacgagc cgggaacaac tcaataccgt aggactgaga
     3481 tatgaaaaaa tttccctccc taaaatacat ccacgctatg atctggatgg ggacgccagc
     3541 atggccaagg cagtgacggg ggttgtgtgc tacgcctgca gaactgccag taccctgctg
     3601 ctctatcaag aattaatgcg aaagggggta cggtggctgg ttgaactggt taaggatgat
     3661 tacaacgaga ccgtccacaa gaagacggag gtagtgatca cactggattt ctgcatcagg
     3721 aacattgaga agactgtgaa agtgtatgag aagttgatga aggtcaacct ggaagccgca
     3781 gagctgggtg agatttcaga catacacacc aagctgctga gactttccag ttctcaggga
     3841 acaatagaaa gcagtcttca ggacatcagc agcaggctgt ctccaggggg cttgctggcc
     3901 gacacctggg cacatcaaga aggcacgcat ccaagagaca ggaatgtaga aaaactgcag
     3961 gtcctgttga actgcatcac agagatttac tatcagttca aaaaagacaa agcagaacgc
     4021 agactagctt ataatgaaga acagatccac aaatttgata agcaaaaatt gtattaccat
     4081 gccacaaaag caatgagcca cttctcagaa gaatgtgtta gaaagtatga agcgtttaaa
     4141 gataagtcgg aagagtggat gagaaagatg cttcatctta ggaagcagct gttatcgcta
     4201 actaatcagt gtttcgatat cgaagaggaa gtgtccaagt atcaagacta tactaacgag
     4261 ttacaagaaa ctctgcctca gaaaatgctc gcagcctccg gcggcgtcaa gcacgccatg
     4321 gccccgatct accccagctc taacacctta gtggagatga ctcttggtat gaagaagtta
     4381 aaggaggaga tggaaggcgt ggttaaggag ctggccgaga acaatcatat tttagaaagg
     4441 tttgggtctt taacaatgga tggtggcctt cgcaatgtgg actgtcttgg atcctaccca
     4501 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct
     4561 tatgatgtcc cagactacgc ataggcggcc gcgaaggatc tgcgatcgct ccggtgcccg
     4621 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     4681 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     4741 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     4801 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
     4861 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
     4921 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
     4981 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     5041 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
     5101 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc
     5161 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     5221 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     5281 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     5341 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     5401 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     5461 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     5521 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     5581 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     5641 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     5701 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc
     5761 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5821 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5881 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5941 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     6001 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     6061 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     6121 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     6181 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     6241 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     6301 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     6361 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     6421 ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta
     6481 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6541 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6601 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6661 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6721 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6781 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6841 catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6901 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6961 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt
     7021 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7081 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7141 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7861 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9181 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     9241 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     9301 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     9361 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     9421 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     9481 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     9541 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     9601 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
