LOCUS       sequence-244573-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of wild-type SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  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 6283)
  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
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..442
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        443..646
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     597..617
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      706..715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2311..3027
                     /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    3149..3270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3186..3205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3240..3259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3277..3732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3626..3645
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3759..3863
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3865..4222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3865..4056
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3888..3908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4073..4208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4257..5051
                     /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             4257..5051
                     /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(4311..4330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4921..4940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5239..5258)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5283..5330
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6148..6167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
       61 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
      121 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      181 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      241 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      301 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      361 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      421 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      481 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      541 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      601 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      661 tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttgccac catggccttc
      721 ttggacaatc caactatcat tctagctcat attcgacagt cacatgtgac cagtgatgac
      781 acgggaatgt gtgagatggt tctcattgat catgatgttg acctagagaa gattcatcct
      841 ccttcaatgc ctggagacag tgggtcagaa attcagggaa gcaatggtga gactcagggc
      901 tatgtatatg ctcagtctgt cgatattacc tcaagttggg actttggtat tagaagacgc
      961 tcaaacacag ctcaaagatt agaacgactc cgaaaagaga gacaaaacca gatcaaatgc
     1021 aaaaatattc agtggaaaga aagaaattct aagcaatcag cccaggagtt aaagtcactg
     1081 tttgaaaaaa aatctctcaa agagaagcct ccaatttctg ggaagcagtc gatattatct
     1141 gtacgcctag aacagtgccc tctgcagctg aataaccctt ttaacgagta ttccaaattt
     1201 gatggcaagg gtcatgtagg tacaacagca accaagaaga tcgatgtcta cctccctctg
     1261 cactcgagcc aggacagact gctgccaatg accgtggtga caatggccag cgccagggtg
     1321 caggacctga tcgggctcat ctgctggcag tatacaagcg aaggacggga gccgaagctc
     1381 aatgacaatg tcagtgccta ctgcctgcat attgctgagg atgatgggga ggtggacacc
     1441 gatttccccc cgctggattc caatgagccc attcataagt ttggcttcag tactttggcc
     1501 ctggttgaaa agtactcatc tcctggtctg acatccaaag agtcactctt tgttcgaata
     1561 aatgctgctc atggattctc ccttattcag gtggacaaca caaaggttac catgaaggaa
     1621 atcttactga aggcagtgaa gcgaagaaaa ggatcccaga aagtttcagg ccctcagtac
     1681 cgcctggaga agcagagcga gcccaatgtc gccgttgacc tggacagcac tttggagagc
     1741 cagagcgcat gggagttctg cctggtccgc gagaacagtt caagggcaga cggggttttt
     1801 gaggaggatt cgcaaattga catagccaca gtacaggata tgcttagcag ccaccattac
     1861 aagtcattca aagtcagcat gatccacaga ctgcgattca caaccgacgt acagctaggt
     1921 atctctggag acaaagtaga gatagaccct gttacgaatc agaaagccag cactaagttt
     1981 tggattaagc agaaacccat ctcaatcgat tccgacctgc tctgtgcctg tgaccttgct
     2041 gaagagaaaa gccccagtca cgcaatattt aaactcacgt atctaagcaa tcacgactat
     2101 aaacacctct actttgaatc ggacgctgct accgtcaatg aaattgtgct caaggttaac
     2161 tacatcctgg aatcgcgagc tagcactgcc cgggctgact actttgctca aaaacaaaga
     2221 aaactgaaca gacgtacgag cttcagcttc cagaaggaga agaaatccgg gcagcaggta
     2281 ccgcgggccc gggatccacc ggtcgccacc atgagcgggg gcgaggagct gttcgccggc
     2341 atcgtgcccg tgctgatcga gctggacggc gacgtgcacg gccacaagtt cagcgtgcgc
     2401 ggcgagggcg agggcgacgc cgactacggc aagctggaga tcaagttcat ctgcaccacc
     2461 ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc tctgctacgg catccagtgc
     2521 ttcgcccgct accccgagca catgaagatg aacgacttct tcaagagcgc catgcccgag
     2581 ggctacatcc aggagcgcac catccagttc caggacgacg gcaagtacaa gacccgcggc
     2641 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg caaggacttc
     2701 aaggaggacg gcaacatcct gggccacaag ctggagtaca gcttcaacag ccacaacgtg
     2761 tacatccgcc ccgacaaggc caacaacggc ctggaggcta acttcaagac ccgccacaac
     2821 atcgagggcg gcggcgtgca gctggccgac cactaccaga ccaacgtgcc cctgggcgac
     2881 ggccccgtgc tgatccccat caaccactac ctgagcactc agaccaagat cagcaaggac
     2941 cgcaacgagg cccgcgacca catggtgctc ctggagtcct tcagcgcctg ctgccacacc
     3001 cacggcatgg acgagctgta caggtaagcg gccgcgactc tagatcataa tcagccatac
     3061 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3121 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3241 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3301 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3421 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3481 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3541 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3601 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3661 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3721 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3781 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3841 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3901 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4261 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4321 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4381 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4441 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4501 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4561 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4621 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4681 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4741 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4801 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4861 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4921 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4981 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5041 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5101 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5161 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5221 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5281 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5341 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5401 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5461 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gcc
//
