LOCUS       sequence-244590-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a.a.116-127 deletion mutant of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  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 6247)
  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 delta116-127
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      675..684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2244..2960
                     /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    3082..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3119..3138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3173..3192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3210..3665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3347..3368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3559..3578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3692..3796
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3798..4155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3798..3989
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3821..3841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4006..4141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4190..4984
                     /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             4190..4984
                     /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(4244..4263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4854..4873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5172..5191)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5216..5263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5592..6180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6081..6100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
      661 tcgagctcaa gcttgccacc atggccttct tggacaatcc aactatcatt ctagctcata
      721 ttcgacagtc acatgtgacc agtgatgaca cgggaatgtg tgagatggtt ctcattgatc
      781 atgatgttga cctagagaag attcatcctc cttcaatgcc tggagacagt gggtcagaaa
      841 ttcagggaag caatggtgag actcagggct atgtatatgc tcagtctgtc gatattacct
      901 caagttggga ctttggtatt agaagacgct caaacacagc tcaaagatta gaacgactcc
      961 gaaaagagag acaaaaccag atcaaatgca aaaatattca gtggaaagaa agaaattcta
     1021 agcaatctct caaagagaag cctccaattt ctgggaagca gtcgatatta tctgtacgcc
     1081 tagaacagtg ccctctgcag ctgaataacc cttttaacga gtattccaaa tttgatggca
     1141 agggtcatgt aggtacaaca gcaaccaaga agatcgatgt ctacctccct ctgcactcga
     1201 gccaggacag actgctgcca atgaccgtgg tgacaatggc cagcgccagg gtgcaggacc
     1261 tgatcgggct catctgctgg cagtatacaa gcgaaggacg ggagccgaag ctcaatgaca
     1321 atgtcagtgc ctactgcctg catattgctg aggatgatgg ggaggtggac accgatttcc
     1381 ccccgctgga ttccaatgag cccattcata agtttggctt cagtactttg gccctggttg
     1441 aaaagtactc atctcctggt ctgacatcca aagagtcact ctttgttcga ataaatgctg
     1501 ctcatggatt ctcccttatt caggtggaca acacaaaggt taccatgaag gaaatcttac
     1561 tgaaggcagt gaagcgaaga aaaggatccc agaaagtttc aggccctcag taccgcctgg
     1621 agaagcagag cgagcccaat gtcgccgttg acctggacag cactttggag agccagagcg
     1681 catgggagtt ctgcctggtc cgcgagaaca gttcaagggc agacggggtt tttgaggagg
     1741 attcgcaaat tgacatagcc acagtacagg atatgcttag cagccaccat tacaagtcat
     1801 tcaaagtcag catgatccac agactgcgat tcacaaccga cgtacagcta ggtatctctg
     1861 gagacaaagt agagatagac cctgttacga atcagaaagc cagcactaag ttttggatta
     1921 agcagaaacc catctcaatc gattccgacc tgctctgtgc ctgtgacctt gctgaagaga
     1981 aaagccccag tcacgcaata tttaaactca cgtatctaag caatcacgac tataaacacc
     2041 tctactttga atcggacgct gctaccgtca atgaaattgt gctcaaggtt aactacatcc
     2101 tggaatcgcg agctagcact gcccgggctg actactttgc tcaaaaacaa agaaaactga
     2161 acagacgtac gagcttcagc ttccagaagg agaagaaatc cgggcagcag gtaccgcggg
     2221 cccgggatcc accggtcgcc accatgagcg ggggcgagga gctgttcgcc ggcatcgtgc
     2281 ccgtgctgat cgagctggac ggcgacgtgc acggccacaa gttcagcgtg cgcggcgagg
     2341 gcgagggcga cgccgactac ggcaagctgg agatcaagtt catctgcacc accggcaagc
     2401 tgcccgtgcc ctggcccacc ctggtgacca ccctctgcta cggcatccag tgcttcgccc
     2461 gctaccccga gcacatgaag atgaacgact tcttcaagag cgccatgccc gagggctaca
     2521 tccaggagcg caccatccag ttccaggacg acggcaagta caagacccgc ggcgaggtga
     2581 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcaaggac ttcaaggagg
     2641 acggcaacat cctgggccac aagctggagt acagcttcaa cagccacaac gtgtacatcc
     2701 gccccgacaa ggccaacaac ggcctggagg ctaacttcaa gacccgccac aacatcgagg
     2761 gcggcggcgt gcagctggcc gaccactacc agaccaacgt gcccctgggc gacggccccg
     2821 tgctgatccc catcaaccac tacctgagca ctcagaccaa gatcagcaag gaccgcaacg
     2881 aggcccgcga ccacatggtg ctcctggagt ccttcagcgc ctgctgccac acccacggca
     2941 tggacgagct gtacaggtaa gcggccgcga ctctagatca taatcagcca taccacattt
     3001 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3061 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3121 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3181 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3241 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3301 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3361 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3421 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3481 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3541 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3601 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3661 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3721 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3781 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3841 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3901 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3961 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4021 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4081 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4141 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4201 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4261 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4321 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4381 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4441 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4501 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4561 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4621 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4681 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4741 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4801 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4861 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4921 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4981 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5041 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5101 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5161 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5221 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5281 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5341 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5401 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5461 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5641 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5701 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5761 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5821 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5881 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5941 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6001 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6061 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6241 ttctttc
//
