LOCUS       sequence-364370-        8240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgYAP-10 sequence: GTGCTGTCCCAGATGAACGTC. Lentiviral mouse U6 (mU6) 
            promoter-driven expression vector that coexpessed Puro-T2A-mCherry 
            from a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8240)
  AUTHORS   Lee JY, Chang JK, Dominguez AA, Lee HP, Nam S, Chang J, Varma S, Qi 
            LS, West RB, Chaudhuri O
  TITLE     YAP-independent mechanotransduction drives breast cancer 
            progression.
  JOURNAL   Nat Commun. 2019 Apr 23;10(1):1848. doi: 10.1038/s41467-019-09755-0.
  PUBMED    31015465
REFERENCE   2  (bases 1 to 8240)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-04-23"; volume: "10(1):1848. doi"; pages: " 
            10.1038/s41467-019-09755-0"
COMMENT     Sequence Label: pSLQ1371-sgYAP-10
FEATURES             Location/Qualifiers
     source          1..8240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..256
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             290..886
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(290..309)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     786..806
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             896..949
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             962..1672
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1107..1125)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1375..1394
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1692..1725)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1781..2369
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1834..1854)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2252..2263)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2372..2388)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2373..2389)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2898..3078
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3140..3728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3220..3239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3899..4690,4691..4759))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4522..4541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4760..4864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4939..4958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5130..5509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5511..5709
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5727..5907
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5954..6079
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6576..6809
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6994..7038
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7336..7453
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(7706..7739)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     7743..7763
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
       61 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      121 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      181 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      241 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcccacca tgaccgagta
      301 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      361 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      421 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      481 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      541 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
      601 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      661 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      721 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
      781 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      841 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggat cgggagaggg
      901 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac cggtcgccac
      961 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     1021 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1081 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1141 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1201 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1261 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1321 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1381 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     1441 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     1501 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     1561 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     1621 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aagaattcgt
     1681 cgagggacct aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt
     1741 ccggttccac taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     1801 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     1861 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     1921 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     1981 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2041 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2101 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2161 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     2221 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2281 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2341 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag
     2401 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     2461 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     2521 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     2581 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     2641 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     2701 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     2761 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     2821 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     2881 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2941 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3001 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3061 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4861 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     4921 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     4981 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     5041 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     5101 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     5161 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5221 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5281 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5341 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5401 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5461 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     5521 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     5581 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     5641 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     5701 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5761 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5821 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5881 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     5941 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     6001 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     6061 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     6121 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     6181 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     6241 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     6301 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     6361 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     6421 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     6481 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     6541 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     6601 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     6661 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     6721 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     6781 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     6841 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     6901 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     6961 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     7021 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     7081 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     7141 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     7201 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     7261 agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc
     7321 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     7381 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     7441 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt tagtaccggg
     7501 cccgctctag agatccgacg cgccatctct aggcccgcgc cggccccctc gcacggactt
     7561 gtgggagaag ctcggctact cccctgcccc ggttaatttg catataatat ttcctagtaa
     7621 ctatagaggc ttaatgtgcg ataaaagaca gataatctgt tctttttaat actagctaca
     7681 ttttacatga taggcttgga tttctataac ttcgtatagc atacattata cgaagttata
     7741 aacagcacaa aaggaaactc accctaactg taaagtaatt gtgtgttttg agactataag
     7801 tatcccttgg agaaccacct tgttggtgct gtcccagatg aacgtcgttt aagagctatg
     7861 ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaacttga aaaagtggca
     7921 ccgagtcggt gctttttttc tcgagtacta ggatccatta ggcggccgcg tggataaccg
     7981 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     8041 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     8101 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     8161 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8221 aagtgtatca tatgccaagt
//
