LOCUS       sequence-244578-        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        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1908..2624
                     /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    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2837..2856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2874..3329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3223..3242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3356..3460
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3462..3819
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3462..3653
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3485..3505)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3670..3805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3854..4648
                     /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             3854..4648
                     /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(3908..3927)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4518..4537
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4836..4855)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4880..4927
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5256..5844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5745..5764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggactcagat ctcgagctca agcttgccac catggccttc ttggacaatc
      661 caactatcat tctagctcat attcgacagt cacatgtgac cagtgatgac acgggaatgt
      721 gtgagatggt tctcattgat catgatgttg acctagagaa gattcatcct ccttcaatgc
      781 ctggagacag tgggtcagaa attcagggaa gcaatggtga gactcagggc tatgtatatg
      841 ctcagtctgt cgatattacc tcaagttggg actttggtat tagaagacgc tcaaacacag
      901 ctcaaagatt agaacgactc cgaaaagaga gacaaaacca gatcaaatgc aaaaatattc
      961 agtggaaaga aagaaattct aagcaatcag cccaggagtt aaagtcactg tttgaaaaaa
     1021 aatctctcaa agagaagcct ccaatttctg ggaagcagtc gatattatct gtacgcctag
     1081 aacagtgccc tctgcagctg aataaccctt ttaacgagta ttccaaattt gatggcaagg
     1141 gtcatgtagg tacaacagca accaagaaga tcgatgtcta cctccctctg cactcgagcc
     1201 aggacagact gctgccaatg accgtggtga caatggccag cgccagggtg caggacctga
     1261 tcgggctcat ctgctggcag tatacaagcg aaggacggga gccgaagctc aatgacaatg
     1321 tcagtgccta ctgcctgcat attgctgagg atgatgggga ggtggacacc gatttccccc
     1381 cgctggattc caatgagccc attcataagt ttggcttcag tactttggcc ctggttgaaa
     1441 agtactcatc tcctggtctg acatccaaag agtcactctt tgttcgaata aatgctgctc
     1501 atggattctc ccttattcag gtggacaaca caaaggttac catgaaggaa atcttactga
     1561 aggcagtgaa gcgaagaaaa ggatcccaga aagtttcagg ccctcagtac cgcctggaga
     1621 agcagagcga gcccaatgtc gccgttgacc tggacagcac tttggagagc cagagcgcat
     1681 gggagttctg cctggtccgc gagaacagtt caagggcaga cggggttttt gaggaggatt
     1741 cgcaaattga catagccaca gtacaggaat cgcgagctag cactgcccgg gctgactact
     1801 ttgctcaaaa acaaagaaaa ctgaacagac gtacgagctt cagcttccag aaggagaaga
     1861 aatccgggca gcaggtaccg cgggcccggg atccaccggt cgccaccatg agcgggggcg
     1921 aggagctgtt cgccggcatc gtgcccgtgc tgatcgagct ggacggcgac gtgcacggcc
     1981 acaagttcag cgtgcgcggc gagggcgagg gcgacgccga ctacggcaag ctggagatca
     2041 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctggtg accaccctct
     2101 gctacggcat ccagtgcttc gcccgctacc ccgagcacat gaagatgaac gacttcttca
     2161 agagcgccat gcccgagggc tacatccagg agcgcaccat ccagttccag gacgacggca
     2221 agtacaagac ccgcggcgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2281 tgaagggcaa ggacttcaag gaggacggca acatcctggg ccacaagctg gagtacagct
     2341 tcaacagcca caacgtgtac atccgccccg acaaggccaa caacggcctg gaggctaact
     2401 tcaagacccg ccacaacatc gagggcggcg gcgtgcagct ggccgaccac taccagacca
     2461 acgtgcccct gggcgacggc cccgtgctga tccccatcaa ccactacctg agcactcaga
     2521 ccaagatcag caaggaccgc aacgaggccc gcgaccacat ggtgctcctg gagtccttca
     2581 gcgcctgctg ccacacccac ggcatggacg agctgtacag gtaagcggcc gcgactctag
     2641 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2701 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2881 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2941 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3001 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3061 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3121 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3181 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3241 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3301 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3421 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3481 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3661 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3721 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3781 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3841 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4021 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4381 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4681 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4861 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4921 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4981 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5041 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5101 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5161 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5221 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5341 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5401 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5941 cgtattaccg
//
