LOCUS       sequence-244583-        6199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a.a.2-29 deletion mutant of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6199)
  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 6199)
  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 delta2-29
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        424..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     578..598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2208..2924
                     /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    3046..3167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3083..3102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3137..3156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3174..3629)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3311..3332)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3523..3542
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3656..3760
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3762..4119
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3762..3953
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3785..3805)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3970..4105
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4154..4948
                     /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             4154..4948
                     /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(4208..4227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4818..4837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5136..5155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5180..5227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5556..6144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6045..6064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      661 cggactcaga tctcgagctc aagcttgcca ccatggttct cattgatcat gatgttgacc
      721 tagagaagat tcatcctcct tcaatgcctg gagacagtgg gtcagaaatt cagggaagca
      781 atggtgagac tcagggctat gtatatgctc agtctgtcga tattacctca agttgggact
      841 ttggtattag aagacgctca aacacagctc aaagattaga acgactccga aaagagagac
      901 aaaaccagat caaatgcaaa aatattcagt ggaaagaaag aaattctaag caatcagccc
      961 aggagttaaa gtcactgttt gaaaaaaaat ctctcaaaga gaagcctcca atttctggga
     1021 agcagtcgat attatctgta cgcctagaac agtgccctct gcagctgaat aaccctttta
     1081 acgagtattc caaatttgat ggcaagggtc atgtaggtac aacagcaacc aagaagatcg
     1141 atgtctacct ccctctgcac tcgagccagg acagactgct gccaatgacc gtggtgacaa
     1201 tggccagcgc cagggtgcag gacctgatcg ggctcatctg ctggcagtat acaagcgaag
     1261 gacgggagcc gaagctcaat gacaatgtca gtgcctactg cctgcatatt gctgaggatg
     1321 atggggaggt ggacaccgat ttccccccgc tggattccaa tgagcccatt cataagtttg
     1381 gcttcagtac tttggccctg gttgaaaagt actcatctcc tggtctgaca tccaaagagt
     1441 cactctttgt tcgaataaat gctgctcatg gattctccct tattcaggtg gacaacacaa
     1501 aggttaccat gaaggaaatc ttactgaagg cagtgaagcg aagaaaagga tcccagaaag
     1561 tttcaggccc tcagtaccgc ctggagaagc agagcgagcc caatgtcgcc gttgacctgg
     1621 acagcacttt ggagagccag agcgcatggg agttctgcct ggtccgcgag aacagttcaa
     1681 gggcagacgg ggtttttgag gaggattcgc aaattgacat agccacagta caggatatgc
     1741 ttagcagcca ccattacaag tcattcaaag tcagcatgat ccacagactg cgattcacaa
     1801 ccgacgtaca gctaggtatc tctggagaca aagtagagat agaccctgtt acgaatcaga
     1861 aagccagcac taagttttgg attaagcaga aacccatctc aatcgattcc gacctgctct
     1921 gtgcctgtga ccttgctgaa gagaaaagcc ccagtcacgc aatatttaaa ctcacgtatc
     1981 taagcaatca cgactataaa cacctctact ttgaatcgga cgctgctacc gtcaatgaaa
     2041 ttgtgctcaa ggttaactac atcctggaat cgcgagctag cactgcccgg gctgactact
     2101 ttgctcaaaa acaaagaaaa ctgaacagac gtacgagctt cagcttccag aaggagaaga
     2161 aatccgggca gcaggtaccg cgggcccggg atccaccggt cgccaccatg agcgggggcg
     2221 aggagctgtt cgccggcatc gtgcccgtgc tgatcgagct ggacggcgac gtgcacggcc
     2281 acaagttcag cgtgcgcggc gagggcgagg gcgacgccga ctacggcaag ctggagatca
     2341 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctggtg accaccctct
     2401 gctacggcat ccagtgcttc gcccgctacc ccgagcacat gaagatgaac gacttcttca
     2461 agagcgccat gcccgagggc tacatccagg agcgcaccat ccagttccag gacgacggca
     2521 agtacaagac ccgcggcgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2581 tgaagggcaa ggacttcaag gaggacggca acatcctggg ccacaagctg gagtacagct
     2641 tcaacagcca caacgtgtac atccgccccg acaaggccaa caacggcctg gaggctaact
     2701 tcaagacccg ccacaacatc gagggcggcg gcgtgcagct ggccgaccac taccagacca
     2761 acgtgcccct gggcgacggc cccgtgctga tccccatcaa ccactacctg agcactcaga
     2821 ccaagatcag caaggaccgc aacgaggccc gcgaccacat ggtgctcctg gagtccttca
     2881 gcgcctgctg ccacacccac ggcatggacg agctgtacag gtaagcggcc gcgactctag
     2941 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3001 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3061 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3121 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3181 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3241 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3301 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3361 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3421 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3481 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3541 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3601 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3661 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3721 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3781 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3841 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3961 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4021 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4081 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4141 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4201 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4261 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4321 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4381 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4441 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4501 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4561 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4621 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4681 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4741 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4801 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4861 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4921 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4981 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5041 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5101 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5161 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5221 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5281 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5341 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5401 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5461 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5881 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5941 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6001 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6061 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6121 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6181 ttttgctggc cttttgctc
//
