LOCUS       sequence-330045-        6183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of mEGFP and YAPS127A- L308/311/325/318E (4LE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6183)
  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 6183)
  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: "Mol Cell. 
            2021 Feb 13. pii: S1097-2765(21)00010-1. doi: 
            10.1016/j.molcel.2021.01.010."
COMMENT     Sequence Label: mEGFP-N1 YAPS127A- L308/311/325/318E (4LE)
FEATURES             Location/Qualifiers
     source          1..6183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2063..2072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2069..2071,2072..2074,2075..2788)
                     /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(2114..2135)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2375..2394)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2722..2743
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2911..3032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2948..2967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3002..3021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3039..3494)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3176..3197)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3388..3407
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3521..3625
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3627..3984
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3627..3818
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3650..3670)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3835..3970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4019..4813
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4019..4813
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4073..4092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4683..4702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5001..5020)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5045..5092
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5421..6009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5910..5929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatggac cccgggcagc agccgccgcc
      601 tcaaccggcc ccccagggcc aagggcagcc gccttcgcag cccccgcagg ggcagggccc
      661 gccgtccgga cccgggcaac cggcacccgc ggcgacccag gcggcgccgc aggcaccccc
      721 cgccgggcat cagatcgtgc acgtccgcgg ggactcggag accgacctgg aggcgctctt
      781 caacgccgtc atgaacccca agacggccaa cgtgccccag accgtgccca tgaggctccg
      841 gaagctgccc gactccttct tcaagccgcc ggagcccaaa tcccactccc gacaggccag
      901 tactgatgca ggcactgcag gagccctgac tccacagcat gttcgagctc atgcctctcc
      961 agcttctctg cagttgggag ctgtttctcc tgggacactg acccccactg gagtagtctc
     1021 tggcccagca gctacaccca cagctcagca tcttcgacag tcttcttttg agatacctga
     1081 tgatgtacct ctgccagcag gttgggagat ggcaaagaca tcttctggtc agagatactt
     1141 cttaaatcac atcgatcaga caacaacatg gcaggacccc aggaaggcca tgctgtccca
     1201 gatgaacgtc acagccccca ccagtccacc agtgcagcag aatatgatga actcggcttc
     1261 aggtcctctt cctgatggat gggaacaagc catgactcag gatggagaaa tttactatat
     1321 aaaccataag aacaagacca cctcttggct agacccaagg cttgaccctc gttttgccat
     1381 gaaccagaga atcagtcaga gtgctccagt gaaacagcca ccacccctgg ctccccagag
     1441 cccacaggga ggcgtcatgg gtggcagcaa ctccaaccag cagcaacaga tgcgagagca
     1501 gcaagagcag atggagaagg agagggagcg gctgaaacag caagaagagc ttcggcagga
     1561 gttagccctg cgtagccagt taccaacact ggagcaggat ggtgggactc aaaatccagt
     1621 gtcttctccc gggatgtctc aggaattgag aacaatgacg accaatagct cagatccttt
     1681 ccttaacagt ggcacctatc actctcgaga tgagagtaca gacagtggac taagcatgag
     1741 cagctacagt gtccctcgaa ccccagatga cttcctgaac agtgtggatg agatggatac
     1801 aggtgatact atcaaccaaa gcaccctgcc ctcacagcag aaccgtttcc cagactacct
     1861 tgaagccatt cctgggacaa atgtggacct tggaacactg gaaggagatg gaatgaacat
     1921 agaaggagag gagctgatgc caagtctgca ggaagctttg agttctgaca tccttaatga
     1981 catggagtct gttttggctg ccaccaagct agataaagaa agctttctta catggttagg
     2041 tggaggcggg tctccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     2101 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2161 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2221 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2281 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2341 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2401 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2461 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2521 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2581 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2641 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga
     2701 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2761 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     2821 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2881 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2941 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3001 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     3061 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3121 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3181 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3241 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3301 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3361 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3421 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3481 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3541 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3601 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3661 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3721 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3781 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3841 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3901 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3961 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4021 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4081 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4141 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4201 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4261 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4321 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4381 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4441 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4501 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4561 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4621 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4681 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4741 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4801 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4861 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4921 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4981 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5041 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5101 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5161 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5221 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6121 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6181 ccg
//
