LOCUS       sequence-329970-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of mEGFP and SAV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  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 5853)
  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-SAV1
FEATURES             Location/Qualifiers
     source          1..5853
                     /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      3083..3092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3089..3091,3092..3094,3095..3808)
                     /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(3089..3091,3092..3094,3095..3805)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3134..3155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3395..3414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3742..3763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3931..4052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3968..3987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4022..4041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4059..4514)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4196..4217)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4408..4427
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4541..4645
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4647..5004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4647..4838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4670..4690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4855..4990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5039..5833
                     /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             5039..5833
                     /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(5093..5112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5703..5722
                     /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 attcatgctg tcccgaaaga aaaccaaaaa cgaagtgtcc aagccggccg aggtgcaggg
     1981 gaagtacgtg aagaaggaga cgtcgcctct gcttcggaat cttatgcctt cattcatccg
     2041 gcatggtcca acaattccaa gacgaactga tatctgtctt ccagattcaa gccctaatgc
     2101 cttttcaact tctggagatg tagtttcaag aaaccagagt ttccttagaa ctccaattca
     2161 aagaacacct catgaaataa tgagaagaga aagcaacaga ttatctgcac cttcttatct
     2221 tgccagaagt ctagcagatg tccctagaga gtatggttct tctcagtcat ttgtaacgga
     2281 agttagtttt gctgttgaaa atggagactc tggttcccga tattattatt cagacaattt
     2341 ttttgatggt cagagaaagc ggccacttgg agatcgtgca catgaagact acagatatta
     2401 tgaatacaac catgatctct tccaaagaat gccacagaat caggggaggc atgcttcagg
     2461 tattgggaga gttgctgcta catctttagg aaatttgact aaccatggtt ctgaagattt
     2521 accccttcct cctggctggt ctgtggactg gacaatgaga gggagaaaat attatataga
     2581 tcataacaca aatacaactc actggagcca tcctcttgag cgagaaggac ttcctcctgg
     2641 atgggaacga gttgagtcat ccgaatttgg aacctattat gtagatcaca caaataagaa
     2701 ggcccaatac aggcatccct gtgctcctag tgtacctcgg tatgatcaac cacctcctgt
     2761 cacataccag ccacagcaaa ctgaaagaaa tcagtccctt ctggtacctg caaatccata
     2821 tcatactgca gaaattcctg actggcttca ggtttacgca cgagcccctg tgaaatatga
     2881 ccacattctg aagtgggaac tcttccagct ggctgacctg gatacatacc agggaatgct
     2941 aaagttgctc ttcatgaaag aattggagca gattgttaaa atgtatgaag catacagaca
     3001 agcccttctt acagagttgg aaaaccgaaa gcagagacag cagtggtatg cccaacaaca
     3061 tggaaaaaat tttccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3121 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3181 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     3301 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3361 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3541 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     3601 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3661 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     3721 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3781 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     3841 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3901 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4021 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     4081 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4141 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4201 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4261 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4321 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4381 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4441 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4501 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4621 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4681 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4741 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4801 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4861 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4921 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4981 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5041 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5101 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5161 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5221 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5281 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5341 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5401 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5461 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5521 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5581 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5641 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5701 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5761 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5821 cgagttcttc tgagcgggac tctggggttc gaa
//
