LOCUS       sequence-244696-        4834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a.a. 1-68 of SIN1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4834)
  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 4834)
  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 Na
FEATURES             Location/Qualifiers
     source          1..4834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      695..704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             923..1639
                     /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    1761..1882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1798..1817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1852..1871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1889..2344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2026..2047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2238..2257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2371..2475
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2500..2520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2544..2754
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2566..2762
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2613..2748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2797..3591
                     /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             2797..3591
                     /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(2851..2870)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3461..3480
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3779..3798)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3823..3870
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4199..4787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4688..4707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
       61 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      661 agcgctaccg gactcagatc tcgagctcaa gcttgccacc atggccttct tggacaatcc
      721 aactatcatt ctagctcata ttcgacagtc acatgtgacc agtgatgaca cgggaatgtg
      781 tgagatggtt ctcattgatc atgatgttga cctagagaag attcatcctc cttcaatgcc
      841 tggagacagt gggtcagaaa ttcagggaag caatggtgag actcagggct atgtatatgc
      901 ccaggatcca ccggtcgcca ccatgagcgg gggcgaggag ctgttcgccg gcatcgtgcc
      961 cgtgctgatc gagctggacg gcgacgtgca cggccacaag ttcagcgtgc gcggcgaggg
     1021 cgagggcgac gccgactacg gcaagctgga gatcaagttc atctgcacca ccggcaagct
     1081 gcccgtgccc tggcccaccc tggtgaccac cctctgctac ggcatccagt gcttcgcccg
     1141 ctaccccgag cacatgaaga tgaacgactt cttcaagagc gccatgcccg agggctacat
     1201 ccaggagcgc accatccagt tccaggacga cggcaagtac aagacccgcg gcgaggtgaa
     1261 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcaaggact tcaaggagga
     1321 cggcaacatc ctgggccaca agctggagta cagcttcaac agccacaacg tgtacatccg
     1381 ccccgacaag gccaacaacg gcctggaggc taacttcaag acccgccaca acatcgaggg
     1441 cggcggcgtg cagctggccg accactacca gaccaacgtg cccctgggcg acggccccgt
     1501 gctgatcccc atcaaccact acctgagcac tcagaccaag atcagcaagg accgcaacga
     1561 ggcccgcgac cacatggtgc tcctggagtc cttcagcgcc tgctgccaca cccacggcat
     1621 ggacgagctg tacaggtaag cggccgcgac tctagatcat aatcagccat accacatttg
     1681 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1741 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1861 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     1921 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1981 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2041 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2101 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2161 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2221 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2281 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2341 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2401 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2461 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     2521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2581 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2641 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2701 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2761 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     2821 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     2881 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     2941 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3001 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3061 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3121 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3181 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3241 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3301 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     3361 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3421 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3481 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3541 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     3601 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3661 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3721 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     3781 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     3841 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     3901 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     3961 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4021 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4081 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4141 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4201 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4261 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4321 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4381 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4441 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4501 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4561 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4621 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4681 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4741 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4801 cggccttttt acggttcctg gccttttgct ggcc
//
