LOCUS       sequence-244589-        6223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a.a.95-114 deletion mutant of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  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 6223)
  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 delta95-114
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        450..653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     604..624
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2258..2974
                     /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    3096..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3133..3152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3187..3206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3224..3679)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3361..3382)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3573..3592
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3706..3810
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3812..4169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3812..4003
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3835..3855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4020..4155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4204..4998
                     /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             4204..4998
                     /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(4258..4277)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4868..4887
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5186..5205)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5230..5277
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5606..6194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6095..6114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
       61 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
      121 ttcatagccc atatatggag ttccgcttta cataacttac ggtaaatggc ccgcctggct
      181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      421 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      481 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      541 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      601 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      661 tgaaccgtca gatccgctag cgctaccgga ctcagatctc gagctcaagc ttgccaccat
      721 ggccttcttg gacaatccaa ctatcattct agctcatatt cgacagtcac atgtgaccag
      781 tgatgacacg ggaatgtgtg agatggttct cattgatcat gatgttgacc tagagaagat
      841 tcatcctcct tcaatgcctg gagacagtgg gtcagaaatt cagggaagca atggtgagac
      901 tcagggctat gtatatgctc agtctgtcga tattacctca agttgggact ttggtattag
      961 aagacgctca aacacagctc aaagattaga acgactccga caatcagccc aggagttaaa
     1021 gtcactgttt gaaaaaaaat ctctcaaaga gaagcctcca atttctggga agcagtcgat
     1081 attatctgta cgcctagaac agtgccctct gcagctgaat aaccctttta acgagtattc
     1141 caaatttgat ggcaagggtc atgtaggtac aacagcaacc aagaagatcg atgtctacct
     1201 ccctctgcac tcgagccagg acagactgct gccaatgacc gtggtgacaa tggccagcgc
     1261 cagggtgcag gacctgatcg ggctcatctg ctggcagtat acaagcgaag gacgggagcc
     1321 gaagctcaat gacaatgtca gtgcctactg cctgcatatt gctgaggatg atggggaggt
     1381 ggacaccgat ttccccccgc tggattccaa tgagcccatt cataagtttg gcttcagtac
     1441 tttggccctg gttgaaaagt actcatctcc tggtctgaca tccaaagagt cactctttgt
     1501 tcgaataaat gctgctcatg gattctccct tattcaggtg gacaacacaa aggttaccat
     1561 gaaggaaatc ttactgaagg cagtgaagcg aagaaaagga tcccagaaag tttcaggccc
     1621 tcagtaccgc ctggagaagc agagcgagcc caatgtcgcc gttgacctgg acagcacttt
     1681 ggagagccag agcgcatggg agttctgcct ggtccgcgag aacagttcaa gggcagacgg
     1741 ggtttttgag gaggattcgc aaattgacat agccacagta caggatatgc ttagcagcca
     1801 ccattacaag tcattcaaag tcagcatgat ccacagactg cgattcacaa ccgacgtaca
     1861 gctaggtatc tctggagaca aagtagagat agaccctgtt acgaatcaga aagccagcac
     1921 taagttttgg attaagcaga aacccatctc aatcgattcc gacctgctct gtgcctgtga
     1981 ccttgctgaa gagaaaagcc ccagtcacgc aatatttaaa ctcacgtatc taagcaatca
     2041 cgactataaa cacctctact ttgaatcgga cgctgctacc gtcaatgaaa ttgtgctcaa
     2101 ggttaactac atcctggaat cgcgagctag cactgcccgg gctgactact ttgctcaaaa
     2161 acaaagaaaa ctgaacagac gtacgagctt cagcttccag aaggagaaga aatccgggca
     2221 gcaggtaccg cgggcccggg atccaccggt cgccaccatg agcgggggcg aggagctgtt
     2281 cgccggcatc gtgcccgtgc tgatcgagct ggacggcgac gtgcacggcc acaagttcag
     2341 cgtgcgcggc gagggcgagg gcgacgccga ctacggcaag ctggagatca agttcatctg
     2401 caccaccggc aagctgcccg tgccctggcc caccctggtg accaccctct gctacggcat
     2461 ccagtgcttc gcccgctacc ccgagcacat gaagatgaac gacttcttca agagcgccat
     2521 gcccgagggc tacatccagg agcgcaccat ccagttccag gacgacggca agtacaagac
     2581 ccgcggcgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcaa
     2641 ggacttcaag gaggacggca acatcctggg ccacaagctg gagtacagct tcaacagcca
     2701 caacgtgtac atccgccccg acaaggccaa caacggcctg gaggctaact tcaagacccg
     2761 ccacaacatc gagggcggcg gcgtgcagct ggccgaccac taccagacca acgtgcccct
     2821 gggcgacggc cccgtgctga tccccatcaa ccactacctg agcactcaga ccaagatcag
     2881 caaggaccgc aacgaggccc gcgaccacat ggtgctcctg gagtccttca gcgcctgctg
     2941 ccacacccac ggcatggacg agctgtacag gtaagcggcc gcgactctag atcataatca
     3001 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3061 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3241 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3301 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3361 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3421 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3481 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3541 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3601 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3661 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3841 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4081 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4141 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4201 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4261 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4321 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4381 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4441 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4501 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4561 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4621 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4681 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4741 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4801 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4861 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4921 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4981 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5041 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5101 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5161 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5221 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5281 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5341 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5401 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gtt
//
