LOCUS       sequence-238753-        5950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Pleckstrin Homology domain deletion mutant of 
            SIN1-GFP (delta376-486).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5950)
  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 5950)
  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 deltaPH
FEATURES             Location/Qualifiers
     source          1..5950
                     /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             1900..2616
                     /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    2738..2859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2829..2848
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2866..3321)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3003..3024)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3215..3234
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3348..3452
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3454..3811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3454..3645
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3477..3497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3662..3797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3846..4640
                     /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             3846..4640
                     /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(3900..3919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4510..4529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4828..4847)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4872..4919
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5248..5836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5737..5756
                     /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 tgggaagcag tcgatattat ctgtacgcct agaacagtgc
     1081 cctctgcagc tgaataaccc ttttaacgag tattccaaat ttgatggcaa gggtcatgta
     1141 ggtacaacag caaccaagaa gatcgatgtc tacctccctc tgcactcgag ccaggacaga
     1201 ctgctgccaa tgaccgtggt gacaatggcc agcgccaggg tgcaggacct gatcgggctc
     1261 atctgctggc agtatacaag cgaaggacgg gagccgaagc tcaatgacaa tgtcagtgcc
     1321 tactgcctgc atattgctga ggatgatggg gaggtggaca ccgatttccc cccgctggat
     1381 tccaatgagc ccattcataa gtttggcttc agtactttgg ccctggttga aaagtactca
     1441 tctcctggtc tgacatccaa agagtcactc tttgttcgaa taaatgctgc tcatggattc
     1501 tcccttattc aggtggacaa cacaaaggtt accatgaagg aaatcttact gaaggcagtg
     1561 aagcgaagaa aaggatccca gaaagtttca ggccctcagt accgcctgga gaagcagagc
     1621 gagcccaatg tcgccgttga cctggacagc actttggaga gccagagcgc atgggagttc
     1681 tgcctggtcc gcgagaacag ttcaagggca gacggggttt ttgaggagga ttcgcaaatt
     1741 gacatagcca cagtacagga atcgcgagct agcactgccc gggctgacta ctttgctcaa
     1801 aaacaaagaa aactgaacag acgtacgagc ttcagcttcc agaaggagaa gaaatccggg
     1861 cagcaggtac cgcgggcccg ggatccaccg gtcgccacca tgagcggggg cgaggagctg
     1921 ttcgccggca tcgtgcccgt gctgatcgag ctggacggcg acgtgcacgg ccacaagttc
     1981 agcgtgcgcg gcgagggcga gggcgacgcc gactacggca agctggagat caagttcatc
     2041 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctgg tgaccaccct ctgctacggc
     2101 atccagtgct tcgcccgcta ccccgagcac atgaagatga acgacttctt caagagcgcc
     2161 atgcccgagg gctacatcca ggagcgcacc atccagttcc aggacgacgg caagtacaag
     2221 acccgcggcg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2281 aaggacttca aggaggacgg caacatcctg ggccacaagc tggagtacag cttcaacagc
     2341 cacaacgtgt acatccgccc cgacaaggcc aacaacggcc tggaggctaa cttcaagacc
     2401 cgccacaaca tcgagggcgg cggcgtgcag ctggccgacc actaccagac caacgtgccc
     2461 ctgggcgacg gccccgtgct gatccccatc aaccactacc tgagcactca gaccaagatc
     2521 agcaaggacc gcaacgaggc ccgcgaccac atggtgctcc tggagtcctt cagcgcctgc
     2581 tgccacaccc acggcatgga cgagctgtac aggtaagcgg ccgcgactct agatcataat
     2641 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2701 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2761 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2821 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     2881 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2941 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3001 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3061 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3121 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3181 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3241 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3301 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3361 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3421 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3721 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3781 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3841 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3901 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3961 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4021 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4081 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4141 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4201 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4261 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4321 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4381 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4441 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4501 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4561 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4621 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4681 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4741 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4801 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4861 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4921 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4981 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5041 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5881 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5941 cgccatgcat
//
