LOCUS       sequence-238752-        5896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Conserved Region In Middle deletion mutant of 
            SIN1-GFP (delta139-267).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  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 5896)
  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 deltaCRIM
FEATURES             Location/Qualifiers
     source          1..5896
                     /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             1846..2562
                     /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    2684..2805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2721..2740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2775..2794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2812..3267)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2949..2970)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3161..3180
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3294..3398
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3400..3757
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3400..3591
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3423..3443)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3608..3743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3792..4586
                     /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             3792..4586
                     /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(3846..3865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4456..4475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4774..4793)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4818..4865
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5194..5782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5683..5702
                     /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 tgctcagtct
      841 gtcgatatta cctcaagttg ggactttggt attagaagac gctcaaacac agctcaaaga
      901 ttagaacgac tccgaaaaga gagacaaaac cagatcaaat gcaaaaatat tcagtggaaa
      961 gaaagaaatt ctaagcaatc agcccaggag ttaaagtcac tgtttgaaaa aaaatctctc
     1021 aaagagaagc ctccaatttc tgggaagtac tcatctcctg gtctgacatc caaagagtca
     1081 ctctttgttc gaataaatgc tgctcatgga ttctccctta ttcaggtgga caacacaaag
     1141 gttaccatga aggaaatctt actgaaggca gtgaagcgaa gaaaaggatc ccagaaagtt
     1201 tcaggccctc agtaccgcct ggagaagcag agcgagccca atgtcgccgt tgacctggac
     1261 agcactttgg agagccagag cgcatgggag ttctgcctgg tccgcgagaa cagttcaagg
     1321 gcagacgggg tttttgagga ggattcgcaa attgacatag ccacagtaca ggatatgctt
     1381 agcagccacc attacaagtc attcaaagtc agcatgatcc acagactgcg attcacaacc
     1441 gacgtacagc taggtatctc tggagacaaa gtagagatag accctgttac gaatcagaaa
     1501 gccagcacta agttttggat taagcagaaa cccatctcaa tcgattccga cctgctctgt
     1561 gcctgtgacc ttgctgaaga gaaaagcccc agtcacgcaa tatttaaact cacgtatcta
     1621 agcaatcacg actataaaca cctctacttt gaatcggacg ctgctaccgt caatgaaatt
     1681 gtgctcaagg ttaactacat cctggaatcg cgagctagca ctgcccgggc tgactacttt
     1741 gctcaaaaac aaagaaaact gaacagacgt acgagcttca gcttccagaa ggagaagaaa
     1801 tccgggcagc aggtaccgcg ggcccgggat ccaccggtcg ccaccatgag cgggggcgag
     1861 gagctgttcg ccggcatcgt gcccgtgctg atcgagctgg acggcgacgt gcacggccac
     1921 aagttcagcg tgcgcggcga gggcgagggc gacgccgact acggcaagct ggagatcaag
     1981 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctggtgac caccctctgc
     2041 tacggcatcc agtgcttcgc ccgctacccc gagcacatga agatgaacga cttcttcaag
     2101 agcgccatgc ccgagggcta catccaggag cgcaccatcc agttccagga cgacggcaag
     2161 tacaagaccc gcggcgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2221 aagggcaagg acttcaagga ggacggcaac atcctgggcc acaagctgga gtacagcttc
     2281 aacagccaca acgtgtacat ccgccccgac aaggccaaca acggcctgga ggctaacttc
     2341 aagacccgcc acaacatcga gggcggcggc gtgcagctgg ccgaccacta ccagaccaac
     2401 gtgcccctgg gcgacggccc cgtgctgatc cccatcaacc actacctgag cactcagacc
     2461 aagatcagca aggaccgcaa cgaggcccgc gaccacatgg tgctcctgga gtccttcagc
     2521 gcctgctgcc acacccacgg catggacgag ctgtacaggt aagcggccgc gactctagat
     2581 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2641 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2821 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2881 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2941 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3001 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3061 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3121 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3181 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3241 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3301 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3361 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3421 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3481 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3601 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3661 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3721 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3781 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3841 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3901 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3961 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4021 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4081 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4141 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4201 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4261 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4321 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4381 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4441 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4501 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4561 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4621 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4741 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4801 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4861 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4921 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4981 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5041 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5101 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5161 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5221 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5281 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5341 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5401 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5461 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5521 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5581 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5641 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5701 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5761 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5821 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5881 tattaccgcc atgcat
//
