LOCUS       V026188                 9653 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026188
VERSION     V026188
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..3018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3055..4578
                     /label="YAP1(NM_001282101)"
                     /note="YAP1(NM_001282101)"
                     /gene="YAP1"
     promoter        4623..4834
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4847..5115
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5142..5741
                     /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    5748..6336
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6410..6643
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6715..6836
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6855..6990
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7028..7044)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7052..7068
                     /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(7076..7106)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7121..7142
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7430..8018)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8189..9049)
                     /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(9050..9154)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9628..9644
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggat
     3061 cccgggcagc agccgccgcc tcaaccggcc ccccagggcc aagggcagcc gccttcgcag
     3121 cccccgcagg ggcagggccc gccgtccgga cccgggcaac cggcacccgc ggcgacccag
     3181 gcggcgccgc aggcaccccc cgccgggcat cagatcgtgc acgtccgcgg ggactcggag
     3241 accgacctgg aggcgctctt caacgccgtc atgaacccca agacggccaa cgtgccccag
     3301 accgtgccca tgaggctccg gaagctgccc gactccttct tcaagccgcc ggagcccaaa
     3361 tcccactccc gacaggccag tactgatgca ggcactgcag gagccctgac tccacagcat
     3421 gttcgagctc attcctctcc agcttctctg cagttgggag ctgtttctcc tgggacactg
     3481 acccccactg gagtagtctc tggcccagca gctacaccca cagctcagca tcttcgacag
     3541 tcttcttttg agatacctga tgatgtacct ctgccagcag gttgggagat ggcaaagaca
     3601 tcttctggtc agagatactt cttaaatcac atcgatcaga caacaacatg gcaggacccc
     3661 aggaaggcca tgctgtccca gatgaacgtc acagccccca ccagtccacc agtgcagcag
     3721 aatatgatga actcggcttc aggtcctctt cctgatggat gggaacaagc catgactcag
     3781 gatggagaaa tttactatat aaaccataag aacaagacca cctcttggct agacccaagg
     3841 cttgaccctc gttttgccat gaaccagaga atcagtcaga gtgctccagt gaaacagcca
     3901 ccacccctgg ctccccagag cccacaggga ggcgtcatgg gtggcagcaa ctccaaccag
     3961 cagcaacaga tgcgactgca gcaactgcag atggagaagg agaggctgcg gctgaaacag
     4021 caagaactgc ttcggcaggt gaggccacag gcaatgcgga atatcaatcc cagcacagca
     4081 aattctccaa aatgtcagga gttagccctg cgtagccagt taccaacact ggagcaggat
     4141 ggtgggactc aaaatccagt gtcttctccc gggatgtctc aggaattgag aacaatgacg
     4201 accaatagct cagatccttt ccttaacagt ggcacctatc actctcgaga tgagagtaca
     4261 gacagtggac taagcatgag cagctacagt gtccctcgaa ccccagatga cttcctgaac
     4321 agtgtggatg agatggatac aggtgatact atcaaccaaa gcaccctgcc ctcacagcag
     4381 aaccgtttcc cagactacct tgaagccatt cctgggacaa atgtggacct tggaacactg
     4441 gaaggagatg gaatgaacat agaaggagag gagctgatgc caagtctgca ggaagctttg
     4501 agttctgaca tccttaatga catggagtct gttttggctg ccaccaagct agataaagaa
     4561 agctttctta catggttata ggcggccgcg aaggatctgc gatcgctccg gtgcccgtca
     4621 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     4681 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     4741 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     4801 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     4861 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     4921 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     4981 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     5041 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     5101 gcgccgttac agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca
     5161 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     5221 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     5281 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5341 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5401 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5461 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     5521 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     5581 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     5641 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     5701 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg
     5761 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5821 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5881 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5941 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6001 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6061 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6121 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6181 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6241 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6301 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     6361 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6421 attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac
     6481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6661 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6721 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6781 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6841 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6961 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt
     7021 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7081 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7141 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7861 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9181 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     9241 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     9301 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     9361 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     9421 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     9481 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     9541 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     9601 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
