LOCUS       sequence-329756-        5014 bp    DNA     circular SYN 14-APR-2025
DEFINITION  HDR template for tagging of endogenous mouse YAP N-terminus with 
            mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  AUTHORS   Yu M, Peng Z, Qin M, Liu Y, Wang J, Zhang C, Lin J, Dong T, Wang L, 
            Li S, Yang Y, Xu S, Guo W, Zhang X, Shi M, Peng H, Luo X, Zhang H, 
            Zhang L, Li Y, Yang XP, Sun S
  TITLE     Interferon-gamma induces tumor resistance to anti-PD-1 immunotherapy
            by promoting YAP phase separation.
  JOURNAL   Mol Cell. 2021 Feb 13. pii: S1097-2765(21)00010-1. doi: 
            10.1016/j.molcel.2021.01.010.
  PUBMED    33606996
REFERENCE   2  (bases 1 to 5014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2021 Feb 13. pii: S1097-2765(21)00010-1. doi: 
            10.1016/j.molcel.2021.01.010."
COMMENT     Sequence Label: pUC19-YAP-mEGFP
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(23..39)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(32..54)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(248..267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     367..389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(427..445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        513..617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(618..686,687..1478)
                     /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     complement(836..855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1649..2237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2138..2157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2391..2408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2525..2546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2561..2566,2567..2584,2585..2591)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2599..2615
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2604..2626
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2623..2639
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2623..2639
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3483..3485,3486..3488,3489..4199)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3528..3549)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3789..3808)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4136..4157
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cccgggtacc gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
       61 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      241 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
      301 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      361 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
      421 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
      481 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      541 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      601 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      661 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      721 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      781 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      841 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      901 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      961 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1021 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1081 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1141 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1201 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1261 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1321 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1381 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1441 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2281 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2341 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2401 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2461 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2521 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2581 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2641 atgattacgc caagcttgca tgcctgcagg tcgactctag agctagcaga ggccctggct
     2701 tggaatccta gctgtctgtg gaagctgtgg cacaaagagt tggaggatat ccttgactac
     2761 agagtaaagc ctgtttaaaa caaaatttaa aaagtaatac tgtttccttt tgtaaggtac
     2821 tcccagtgca actcctggtt tccaagcttc actttgaaat catttgcacc cgccctcaag
     2881 tgttttgttg tttcttcttc aaacttggtg gtaaatggaa tcaatgttaa aacgcggacc
     2941 ggattaacct tagtgcgggt gaacagtgtg gcctttgcat ccaccctacc aagccggttg
     3001 tcccgattcc tcgcggcctt ggcgttgcgc gcgggtccgg caggatctgc gcccgcagcc
     3061 gagcggctcg gggccggccg gtgggtgcgc acgcagcgcg gtcgcccccg ggtcacgtgc
     3121 ggccgggagt ctgcaggaat gtagcgacgc gcagggaaag tttccccggg ggccgcgggc
     3181 cgcggattcg gtgcgggcgg gcgcgcagag gaaggaagag cgtcgaggag gcggagcccg
     3241 cgaggcgcgc gggcgggcgc tcctcgcaac tttttagttt gggcgtctgg agccaaagtt
     3301 tctgtctcag ttgggacgcc gccgcggccg ggggcaaaga aagggaggaa ggaaggagct
     3361 cgcggagggg aggggaggag aggggaggcg gcctcgggca aggagtgcag ggcgatgcgg
     3421 gcgcgcgtcg cagccccccg aacctgagcg cagtgccccg agcgtcgaac gaggccgcag
     3481 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3541 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3601 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3661 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3721 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3781 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3841 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3901 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3961 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4021 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4081 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     4141 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     4201 gtggaggcgg gtctatggag cccgcgcaac agccgccgcc ccagccggcc ccgcaaggcc
     4261 ccgcgccgcc gtccgtgtct ccggccggga cccccgcggc cccgcccgca cccccggccg
     4321 gccaccaggt cgtgcacgtc cgcggggact cggagaccga cttggaggcg ctcttcaatg
     4381 ccgtcatgaa ccccaagacg gccaacgtgc ctcagaccgt gcccatgcgg cttcgcaagc
     4441 tgcccgactc cttcttcaag ccgcctgagc ccaagtccca ctcgcgacag gtaagggtat
     4501 cccgggggcg cggggccgcg tgggggtgcg gtgctggggg ttgttttcgc ggagttggcc
     4561 gcaggggggg tgcacaatgc gcgcgcctcc gctccgcacc ttcgctctga cgagggggag
     4621 gggaggaggg ggagggggcc ggcgtcttgt gcgggacccc agccgggggt acccgatctc
     4681 cccttcccgc gccactgcag tctccggtgg agctgtgtta actgccctct tgttttcgcg
     4741 tgccggtgag gggtgggggt tgaactggtc cattgagttt ctggttcctt ctcccggtcc
     4801 cctccccctc cctctcccga aagcttttct gttctcgcgg cccaggcgct ctccggatgc
     4861 agggttcggg ctccccagcc cgagcgaagc tccctcgacc ccccactcgc ctccgccgtg
     4921 ccctggaact cgcgcgggcg ggatcctttc acactcggtg ggctcacgtg tccttttcct
     4981 aaatcgtaac ctattattta ctgttcggcc cgaa
//
