LOCUS       sequence-244575-        5875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of N-terminal Conserved Domain deletion mutant of 
            SIN1-GFP (delta2-137).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5875)
  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 5875)
  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 deltaNCD
FEATURES             Location/Qualifiers
     source          1..5875
                     /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"
     CDS             1872..2588
                     /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    2710..2831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2747..2766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2801..2820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2838..3293)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2975..2996)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3187..3206
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3320..3424
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3426..3783
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3426..3617
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3449..3469)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3634..3769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3818..4612
                     /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             3818..4612
                     /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(3872..3891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4482..4501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4800..4819)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4844..4891
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5220..5808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5709..5728
                     /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 atgaagcagt cgatattatc tgtacgccta gaacagtgcc
      721 ctctgcagct gaataaccct tttaacgagt attccaaatt tgatggcaag ggtcatgtag
      781 gtacaacagc aaccaagaag atcgatgtct acctccctct gcactcgagc caggacagac
      841 tgctgccaat gaccgtggtg acaatggcca gcgccagggt gcaggacctg atcgggctca
      901 tctgctggca gtatacaagc gaaggacggg agccgaagct caatgacaat gtcagtgcct
      961 actgcctgca tattgctgag gatgatgggg aggtggacac cgatttcccc ccgctggatt
     1021 ccaatgagcc cattcataag tttggcttca gtactttggc cctggttgaa aagtactcat
     1081 ctcctggtct gacatccaaa gagtcactct ttgttcgaat aaatgctgct catggattct
     1141 cccttattca ggtggacaac acaaaggtta ccatgaagga aatcttactg aaggcagtga
     1201 agcgaagaaa aggatcccag aaagtttcag gccctcagta ccgcctggag aagcagagcg
     1261 agcccaatgt cgccgttgac ctggacagca ctttggagag ccagagcgca tgggagttct
     1321 gcctggtccg cgagaacagt tcaagggcag acggggtttt tgaggaggat tcgcaaattg
     1381 acatagccac agtacaggat atgcttagca gccaccatta caagtcattc aaagtcagca
     1441 tgatccacag actgcgattc acaaccgacg tacagctagg tatctctgga gacaaagtag
     1501 agatagaccc tgttacgaat cagaaagcca gcactaagtt ttggattaag cagaaaccca
     1561 tctcaatcga ttccgacctg ctctgtgcct gtgaccttgc tgaagagaaa agccccagtc
     1621 acgcaatatt taaactcacg tatctaagca atcacgacta taaacacctc tactttgaat
     1681 cggacgctgc taccgtcaat gaaattgtgc tcaaggttaa ctacatcctg gaatcgcgag
     1741 ctagcactgc ccgggctgac tactttgctc aaaaacaaag aaaactgaac agacgtacga
     1801 gcttcagctt ccagaaggag aagaaatccg ggcagcaggt accgcgggcc cgggatccac
     1861 cggtcgccac catgagcggg ggcgaggagc tgttcgccgg catcgtgccc gtgctgatcg
     1921 agctggacgg cgacgtgcac ggccacaagt tcagcgtgcg cggcgagggc gagggcgacg
     1981 ccgactacgg caagctggag atcaagttca tctgcaccac cggcaagctg cccgtgccct
     2041 ggcccaccct ggtgaccacc ctctgctacg gcatccagtg cttcgcccgc taccccgagc
     2101 acatgaagat gaacgacttc ttcaagagcg ccatgcccga gggctacatc caggagcgca
     2161 ccatccagtt ccaggacgac ggcaagtaca agacccgcgg cgaggtgaag ttcgagggcg
     2221 acaccctggt gaaccgcatc gagctgaagg gcaaggactt caaggaggac ggcaacatcc
     2281 tgggccacaa gctggagtac agcttcaaca gccacaacgt gtacatccgc cccgacaagg
     2341 ccaacaacgg cctggaggct aacttcaaga cccgccacaa catcgagggc ggcggcgtgc
     2401 agctggccga ccactaccag accaacgtgc ccctgggcga cggccccgtg ctgatcccca
     2461 tcaaccacta cctgagcact cagaccaaga tcagcaagga ccgcaacgag gcccgcgacc
     2521 acatggtgct cctggagtcc ttcagcgcct gctgccacac ccacggcatg gacgagctgt
     2581 acaggtaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     2641 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2701 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2761 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2821 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2881 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2941 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3001 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3061 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3121 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3181 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3241 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3301 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3361 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3421 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3541 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3601 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3661 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3721 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3781 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3841 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3901 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3961 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4021 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4081 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4141 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4201 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4261 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4321 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4381 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4441 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4501 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4561 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4621 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4681 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4741 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4801 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4861 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4921 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4981 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5041 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5341 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttc
//
