LOCUS       sequence-244592-        5119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of NCD (a.a. 1-139) of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5119)
  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 5119)
  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 NCD
FEATURES             Location/Qualifiers
     source          1..5119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1076..1792
                     /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    1914..2035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1951..1970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2005..2024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2042..2497)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2179..2200)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2391..2410
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2524..2628
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2630..2987
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2630..2821
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2653..2673)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2838..2973
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3022..3816
                     /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             3022..3816
                     /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(3076..3095)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3686..3705
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4004..4023)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4048..4095
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4424..5012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4913..4932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttgccacc atggccttct tggacaatcc
      661 aactatcatt ctagctcata ttcgacagtc acatgtgacc agtgatgaca cgggaatgtg
      721 tgagatggtt ctcattgatc atgatgttga cctagagaag attcatcctc cttcaatgcc
      781 tggagacagt gggtcagaaa ttcagggaag caatggtgag actcagggct atgtatatgc
      841 tcagtctgtc gatattacct caagttggga ctttggtatt agaagacgct caaacacagc
      901 tcaaagatta gaacgactcc gaaaagagag acaaaaccag atcaaatgca aaaatattca
      961 gtggaaagaa agaaattcta agcaatcagc ccaggagtta aagtcactgt ttgaaaaaaa
     1021 atctctcaaa gagaagcctc caatttctgg gaagcaggat ccaccggtcg ccaccatgag
     1081 cgggggcgag gagctgttcg ccggcatcgt gcccgtgctg atcgagctgg acggcgacgt
     1141 gcacggccac aagttcagcg tgcgcggcga gggcgagggc gacgccgact acggcaagct
     1201 ggagatcaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctggtgac
     1261 caccctctgc tacggcatcc agtgcttcgc ccgctacccc gagcacatga agatgaacga
     1321 cttcttcaag agcgccatgc ccgagggcta catccaggag cgcaccatcc agttccagga
     1381 cgacggcaag tacaagaccc gcggcgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1441 catcgagctg aagggcaagg acttcaagga ggacggcaac atcctgggcc acaagctgga
     1501 gtacagcttc aacagccaca acgtgtacat ccgccccgac aaggccaaca acggcctgga
     1561 ggctaacttc aagacccgcc acaacatcga gggcggcggc gtgcagctgg ccgaccacta
     1621 ccagaccaac gtgcccctgg gcgacggccc cgtgctgatc cccatcaacc actacctgag
     1681 cactcagacc aagatcagca aggaccgcaa cgaggcccgc gaccacatgg tgctcctgga
     1741 gtccttcagc gcctgctgcc acacccacgg catggacgag ctgtacaggt aagcggccgc
     1801 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1861 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1921 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1981 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2041 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2101 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2161 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2221 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2281 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2341 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2401 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2461 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2521 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2581 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2641 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2821 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2881 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2941 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3001 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3061 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3121 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3181 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3241 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3301 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3361 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3421 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3481 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3541 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3601 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3661 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3721 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3781 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3841 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     3901 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3961 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4021 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4081 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4141 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4201 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4261 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4321 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4381 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4441 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4501 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4561 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4621 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4681 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4741 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4801 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4861 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4921 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4981 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5041 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5101 tggataaccg tattaccgc
//
