LOCUS       sequence-234648-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgYAP-2 sequence: GAGATGACTTCCTGAACAGTG. 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 8308)
  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 8308)
  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-2
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             543..1139
                     /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(543..562)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1039..1059
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1149..1202
                     /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      1209..1218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1215..1925
                     /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(1360..1378)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1628..1647
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1945..1978)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2034..2622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2087..2107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2505..2516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2625..2641)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2626..2642)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3151..3331
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3393..3981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3473..3492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4152..4943,4944..5012))
                     /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     4775..4794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5013..5117)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5192..5211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5383..5762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5764..5962
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5980..6160
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6207..6332
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6829..7062
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7247..7291
                     /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    7589..7706
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(7959..7992)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     7996..8016
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagccca
      541 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
      601 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
      661 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
      721 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
      781 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
      841 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
      901 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
      961 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     1021 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     1081 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgccg
     1141 gatcgggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     1201 caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1261 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1321 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1381 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1441 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1501 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1561 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1621 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     1681 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1741 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1801 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1861 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1921 agtaagaatt cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac
     1981 atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac
     2041 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2101 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2161 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2221 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2281 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2341 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2401 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     2461 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2521 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2581 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     2641 gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     2701 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     2761 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     2821 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     2881 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     2941 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     3001 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     3061 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
     3121 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     3181 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3241 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3301 cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc
     3361 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3421 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3481 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3541 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3601 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3661 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3721 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3781 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3841 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3901 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3961 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4021 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4081 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4141 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4201 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4261 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4321 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4381 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4441 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4501 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4561 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4621 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4681 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4741 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4801 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4861 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4921 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4981 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5041 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5101 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5161 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5221 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     5281 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     5341 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     5401 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     5461 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     5521 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     5581 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5641 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5701 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5761 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     5821 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5881 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5941 ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag
     6001 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6061 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     6121 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     6181 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     6241 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     6301 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     6361 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     6421 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     6481 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     6541 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     6601 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     6661 cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     6721 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     6781 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     6841 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     6901 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     6961 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     7021 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     7081 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     7141 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     7201 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     7261 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     7321 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     7381 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     7441 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     7501 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
     7561 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     7621 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     7681 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     7741 ggttagtacc gggcccgctc tagagatccg acgcgccatc tctaggcccg cgccggcccc
     7801 ctcgcacgga cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa
     7861 tatttcctag taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt
     7921 aatactagct acattttaca tgataggctt ggatttctat aacttcgtat agcatacatt
     7981 atacgaagtt ataaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt
     8041 ttgagactat aagtatccct tggagaacca ccttgttgga gatgacttcc tgaacagtgg
     8101 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
     8161 tgaaaaagtg gcaccgagtc ggtgcttttt ttctcgagta ctaggatcca ttaggcggcc
     8221 gcgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     8281 ttagttcata gcccatatat ggagttcc
//
