LOCUS       sequence-244588-        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        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     522..542
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      631..640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2865
                     /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    2987..3108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3024..3043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3078..3097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3115..3570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3252..3273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3464..3483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3597..3701
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3703..4060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3703..3894
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3726..3746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4095..4889
                     /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             4095..4889
                     /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(4149..4168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4759..4778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5077..5096)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5121..5168
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5497..6085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5986..6005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      601 ctaccggact cagatctcga gctcaagctt gccaccatgg ccttcttgga caatccaact
      661 atcattctag ctcatattcg acagtcacat gtgaccagtg atgacacggg aatgtgtgag
      721 atggttctca ttgatcatga tgttgaccta gagaagattc atcctccttc aatgcctgga
      781 gacagtgggt cagaaattca gggaagcaat ggtgagactc agggctatgt atatgctcag
      841 caaaaccaga tcaaatgcaa aaatattcag tggaaagaaa gaaattctaa gcaatcagcc
      901 caggagttaa agtcactgtt tgaaaaaaaa tctctcaaag agaagcctcc aatttctggg
      961 aagcagtcga tattatctgt acgcctagaa cagtgccctc tgcagctgaa taaccctttt
     1021 aacgagtatt ccaaatttga tggcaagggt catgtaggta caacagcaac caagaagatc
     1081 gatgtctacc tccctctgca ctcgagccag gacagactgc tgccaatgac cgtggtgaca
     1141 atggccagcg ccagggtgca ggacctgatc gggctcatct gctggcagta tacaagcgaa
     1201 ggacgggagc cgaagctcaa tgacaatgtc agtgcctact gcctgcatat tgctgaggat
     1261 gatggggagg tggacaccga tttccccccg ctggattcca atgagcccat tcataagttt
     1321 ggcttcagta ctttggccct ggttgaaaag tactcatctc ctggtctgac atccaaagag
     1381 tcactctttg ttcgaataaa tgctgctcat ggattctccc ttattcaggt ggacaacaca
     1441 aaggttacca tgaaggaaat cttactgaag gcagtgaagc gaagaaaagg atcccagaaa
     1501 gtttcaggcc ctcagtaccg cctggagaag cagagcgagc ccaatgtcgc cgttgacctg
     1561 gacagcactt tggagagcca gagcgcatgg gagttctgcc tggtccgcga gaacagttca
     1621 agggcagacg gggtttttga ggaggattcg caaattgaca tagccacagt acaggatatg
     1681 cttagcagcc accattacaa gtcattcaaa gtcagcatga tccacagact gcgattcaca
     1741 accgacgtac agctaggtat ctctggagac aaagtagaga tagaccctgt tacgaatcag
     1801 aaagccagca ctaagttttg gattaagcag aaacccatct caatcgattc cgacctgctc
     1861 tgtgcctgtg accttgctga agagaaaagc cccagtcacg caatatttaa actcacgtat
     1921 ctaagcaatc acgactataa acacctctac tttgaatcgg acgctgctac cgtcaatgaa
     1981 attgtgctca aggttaacta catcctggaa tcgcgagcta gcactgcccg ggctgactac
     2041 tttgctcaaa aacaaagaaa actgaacaga cgtacgagct tcagcttcca gaaggagaag
     2101 aaatccgggc agcaggtacc gcgggcccgg gatccaccgg tcgccaccat gagcgggggc
     2161 gaggagctgt tcgccggcat cgtgcccgtg ctgatcgagc tggacggcga cgtgcacggc
     2221 cacaagttca gcgtgcgcgg cgagggcgag ggcgacgccg actacggcaa gctggagatc
     2281 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt gaccaccctc
     2341 tgctacggca tccagtgctt cgcccgctac cccgagcaca tgaagatgaa cgacttcttc
     2401 aagagcgcca tgcccgaggg ctacatccag gagcgcacca tccagttcca ggacgacggc
     2461 aagtacaaga cccgcggcga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2521 ctgaagggca aggacttcaa ggaggacggc aacatcctgg gccacaagct ggagtacagc
     2581 ttcaacagcc acaacgtgta catccgcccc gacaaggcca acaacggcct ggaggctaac
     2641 ttcaagaccc gccacaacat cgagggcggc ggcgtgcagc tggccgacca ctaccagacc
     2701 aacgtgcccc tgggcgacgg ccccgtgctg atccccatca accactacct gagcactcag
     2761 accaagatca gcaaggaccg caacgaggcc cgcgaccaca tggtgctcct ggagtccttc
     2821 agcgcctgct gccacaccca cggcatggac gagctgtaca ggtaagcggc cgcgactcta
     2881 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2941 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3001 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3061 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3121 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3181 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3241 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3301 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3361 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3421 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3481 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3541 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3601 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3661 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3721 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3841 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3961 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4081 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4141 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4201 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4261 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4321 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4381 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4441 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4501 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4561 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4621 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4681 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4741 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4801 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4861 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4921 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4981 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5041 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5101 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5161 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5221 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5281 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5341 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5401 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5461 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5521 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5581 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5641 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5701 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5761 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5821 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5881 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5941 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6001 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6061 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6121 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6181 ccgtattacc gccatg
//
