LOCUS       sequence-329964-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of mEGFP and NF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  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 6477)
  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-NF2
FEATURES             Location/Qualifiers
     source          1..6477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(168..187)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    212..259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      588..1176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1077..1096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1351..1654
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1655..1858
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1809..1829
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3707..3716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3713..3715,3716..3718,3719..4432)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3713..3715,3716..3718,3719..4429)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3758..3779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4019..4038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4366..4387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4555..4676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4592..4611)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4646..4665
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4683..5138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4820..4841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5032..5051
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5165..5269
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5271..5628
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5271..5462
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5294..5314)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5479..5614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5663..6457
                     /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             5663..6457
                     /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(5717..5736)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6327..6346
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
       61 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
      121 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
      181 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
      241 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
      301 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
      361 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
      421 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
      481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
      541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
      601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      721 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      961 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1261 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     1321 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1381 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1441 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1501 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1561 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1621 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1681 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1741 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1801 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     1861 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga
     1921 attcatggcc ggagccatcg cttctcgcat gagcttcagc tcactcaaga ggaagcagcc
     1981 caagacattc acggtgcgga tcgtcaccat ggacgccgag atggagttca actgcgagat
     2041 gaaatggaag gggaaggacc tgtttgattt ggtgtgccgg acactggggc ttcgggaaac
     2101 ctggttcttt ggactgcagt atacaatcaa ggacacggtg gcctggctca aaatggacaa
     2161 gaaggtgttg gatcatgatg tttcgaagga agaaccagtt acctttcact tcctggccaa
     2221 attttatcct gaaaatgctg aggaggagct agttcaagag atcacgcaac acttattttt
     2281 cttacaggtg aagaagcaga ttttggatga aaaggtctac tgccctcccg aggcgtccgt
     2341 gctcttggcg tcatatgctg tccaggccaa gtatggcgac tatgacccct ctgtgcacaa
     2401 gcggggattt ttagcccaag aggaattgct cccgaaaagg gtgataaatc tctatcagat
     2461 gactccggaa atgtgggagg agagaattac ggcttggtat gcggagcacc ggggcagagc
     2521 cagggatgaa gctgaaatgg agtatttgaa gatagctcag gacctggaga tgtatggtgt
     2581 gaactacttt acaatccgga ataaaaaggg cacagagttg ctgcttggag tggatgctct
     2641 tgggcttcat atctatgacc ctgagaacag gctgaccccc aagatctcct tcccatggaa
     2701 tgaaatccga aacatctcct acagcgacaa ggagtttact attaaaccac tggataagaa
     2761 aattgatgtc ttcaaattta actcctcaaa gcttcgtgtt aataagctga ttcttcagct
     2821 atgtattggg aaccatgacc tatttatgag gcgacggaaa gctgactctt tagaagttca
     2881 gcagatgaaa gcccaggcca gggaagagaa ggctagaaag cagatggaaa ggcagcggct
     2941 ggctcgagag aagcagatgc gggaggaggc cgagcgtaca agagatgagt tagagaggag
     3001 gctcctgcag atgaaagaag aagcaacgat ggccaatgaa gctctgatgc gctctgagga
     3061 gacagctgat ctgttggctg aaaaggctca gatcacagag gaggaggcca agcttttggc
     3121 acaaaaggct gcagaggctg agcaagagat gcagcgaatc aaggccacgg ccattcggac
     3181 agaggaggag aagcgcctga tggagcagaa ggtgctggag gctgaagtgc tggcattgaa
     3241 gatggctgag gagtcagaga ggagggccaa ggaggctgat cagttaaagc aagacttgca
     3301 agaagccaga gaagcagagc gaagagccaa gcagaagctc ttagaaatcg ccaccaagcc
     3361 cacctatcca cccatgaacc caattccacc accactgcct cctgacatac cgagcttcga
     3421 cattattgct gacagcttgt catttgactt caaggatacg gacatgaagc gactttccat
     3481 ggagatagag aaagaaaaag tggagtacat ggagaagagc aagcacctgc aggagcagct
     3541 caacgagctc aagacggaga tcgaggcctt gaaactcaaa gagcgggaga cggccttgga
     3601 cgtcctacac agcgagagct cagacagagg cggccccagc agcaagcata ataccattaa
     3661 aaagcctcaa gcccaaggca gaagacctat ctgcattcca ccggtcgcca ccatggtgag
     3721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3901 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     4081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     4141 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4201 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     4441 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4501 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4561 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4621 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     4681 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4741 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4801 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4861 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4921 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4981 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5041 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5101 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5161 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5221 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5281 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5401 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5461 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5581 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     5641 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5701 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5761 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5821 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     5881 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5941 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     6001 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     6061 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     6121 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     6181 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     6241 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     6301 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     6361 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     6421 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaa
//
