LOCUS       sequence-238755-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a.a.69-97 deletion mutant of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  AUTHORS   Hwang Y, Kim LC, Song W, Edwards DN, Cook RS, Chen J
  TITLE     Disruption of the Scaffolding Function of mLST8 Selectively Inhibits
            mTORC2 Assembly and Function and Suppresses mTORC2-Dependent Tumor 
            Growth In Vivo.
  JOURNAL   Cancer Res. 2019 Jul 1;79(13):3178-3184. doi: 
            10.1158/0008-5472.CAN-18-3658. Epub 2019 May 13.
  PUBMED    31085701
REFERENCE   2  (bases 1 to 6196)
  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"; doi: 
            "10.1158/0008-5472.CAN-18-3658"; journalName: "Cancer Res"; date: 
            "2019-07-1- 1"; volume: "79"; issue: "13"; pages: "3178-3184"
COMMENT     Sequence Label: pTagGFP2-N SIN1 delta69-97
FEATURES             Location/Qualifiers
     source          1..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2146..2862
                     /codon_start=1
                     /product="monomeric green fluorescent protein, also known 
                     as mTagGFP"
                     /label=TagGFP2
                     /note="mammalian codon-optimized"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     polyA_signal    2984..3105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3021..3040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3075..3094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3112..3567)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3249..3270)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3461..3480
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3594..3698
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3700..4057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3700..3891
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3723..3743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3908..4043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4092..4886
                     /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             4092..4886
                     /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(4146..4165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4756..4775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5074..5093)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5118..5165
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5494..6082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5983..6002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc accatggcct tcttggacaa tccaactatc
      661 attctagctc atattcgaca gtcacatgtg accagtgatg acacgggaat gtgtgagatg
      721 gttctcattg atcatgatgt tgacctagag aagattcatc ctccttcaat gcctggagac
      781 agtgggtcag aaattcaggg aagcaatggt gagactcagg gctatgtata tgctcagcaa
      841 aaccagatca aatgcaaaaa tattcagtgg aaagaaagaa attctaagca atcagcccag
      901 gagttaaagt cactgtttga aaaaaaatct ctcaaagaga agcctccaat ttctgggaag
      961 cagtcgatat tatctgtacg cctagaacag tgccctctgc agctgaataa cccttttaac
     1021 gagtattcca aatttgatgg caagggtcat gtaggtacaa cagcaaccaa gaagatcgat
     1081 gtctacctcc ctctgcactc gagccaggac agactgctgc caatgaccgt ggtgacaatg
     1141 gccagcgcca gggtgcagga cctgatcggg ctcatctgct ggcagtatac aagcgaagga
     1201 cgggagccga agctcaatga caatgtcagt gcctactgcc tgcatattgc tgaggatgat
     1261 ggggaggtgg acaccgattt ccccccgctg gattccaatg agcccattca taagtttggc
     1321 ttcagtactt tggccctggt tgaaaagtac tcatctcctg gtctgacatc caaagagtca
     1381 ctctttgttc gaataaatgc tgctcatgga ttctccctta ttcaggtgga caacacaaag
     1441 gttaccatga aggaaatctt actgaaggca gtgaagcgaa gaaaaggatc ccagaaagtt
     1501 tcaggccctc agtaccgcct ggagaagcag agcgagccca atgtcgccgt tgacctggac
     1561 agcactttgg agagccagag cgcatgggag ttctgcctgg tccgcgagaa cagttcaagg
     1621 gcagacgggg tttttgagga ggattcgcaa attgacatag ccacagtaca ggatatgctt
     1681 agcagccacc attacaagtc attcaaagtc agcatgatcc acagactgcg attcacaacc
     1741 gacgtacagc taggtatctc tggagacaaa gtagagatag accctgttac gaatcagaaa
     1801 gccagcacta agttttggat taagcagaaa cccatctcaa tcgattccga cctgctctgt
     1861 gcctgtgacc ttgctgaaga gaaaagcccc agtcacgcaa tatttaaact cacgtatcta
     1921 agcaatcacg actataaaca cctctacttt gaatcggacg ctgctaccgt caatgaaatt
     1981 gtgctcaagg ttaactacat cctggaatcg cgagctagca ctgcccgggc tgactacttt
     2041 gctcaaaaac aaagaaaact gaacagacgt acgagcttca gcttccagaa ggagaagaaa
     2101 tccgggcagc aggtaccgcg ggcccgggat ccaccggtcg ccaccatgag cgggggcgag
     2161 gagctgttcg ccggcatcgt gcccgtgctg atcgagctgg acggcgacgt gcacggccac
     2221 aagttcagcg tgcgcggcga gggcgagggc gacgccgact acggcaagct ggagatcaag
     2281 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctggtgac caccctctgc
     2341 tacggcatcc agtgcttcgc ccgctacccc gagcacatga agatgaacga cttcttcaag
     2401 agcgccatgc ccgagggcta catccaggag cgcaccatcc agttccagga cgacggcaag
     2461 tacaagaccc gcggcgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2521 aagggcaagg acttcaagga ggacggcaac atcctgggcc acaagctgga gtacagcttc
     2581 aacagccaca acgtgtacat ccgccccgac aaggccaaca acggcctgga ggctaacttc
     2641 aagacccgcc acaacatcga gggcggcggc gtgcagctgg ccgaccacta ccagaccaac
     2701 gtgcccctgg gcgacggccc cgtgctgatc cccatcaacc actacctgag cactcagacc
     2761 aagatcagca aggaccgcaa cgaggcccgc gaccacatgg tgctcctgga gtccttcagc
     2821 gcctgctgcc acacccacgg catggacgag ctgtacaggt aagcggccgc gactctagat
     2881 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2941 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3121 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3181 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3241 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3301 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3361 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3421 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3481 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3541 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3601 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3661 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4021 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5041 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5101 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5161 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5221 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5281 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5641 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6181 tattaccgcc atgcat
//
