LOCUS       sequence-238750-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      628..637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2233..2949
                     /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    3071..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3108..3127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3162..3181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3199..3654)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3336..3357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3548..3567
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3681..3785
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3787..4144
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3787..3978
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3810..3830)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3995..4130
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4179..4973
                     /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             4179..4973
                     /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(4233..4252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4843..4862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5161..5180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5205..5252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5581..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6070..6089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc accatggcct tcttggacaa tccaactatc
      661 attctagctc atattcgaca gtcacatgtg accagtgatg acacgggaat gtgtgagatg
      721 gttctcattg atcatgatgt tgacctagag aagattcatc ctccttcaat gcctggagac
      781 agtgggtcag aaattcaggg aagcaatggt gagactcagg gctatgtata tgctcagtct
      841 gtcgatatta cctcaagttg ggactttggt attagaagac gctcaaacac agctcaaaga
      901 ttagaacgac tccgaaaaga gagacaaaac cagatcaaat gcaaaaatat tcagtggaaa
      961 gaaagaaatt ctaagcaatc agcccaggag ttaaagtcac tgtttgaaaa aaaatctctc
     1021 aaagagaagc ctccaatttc tgggaagcag tcgatattat ctgtacgcct agaacagtgc
     1081 cctctgcagc tgaataaccc ttttaacgag tattccaaat ttgatggcaa gggtcatgta
     1141 ggtacaacag caaccaagaa gatcgatgtc tacctccctc tgcactcgag ccaggacaga
     1201 ctgctgccaa tgaccgtggt gacaatggcc agcgccaggg tgcaggacct gatcgggctc
     1261 atctgctggc agtatacaag cgaaggacgg gagccgaagc tcaatgacaa tgtcagtgcc
     1321 tactgcctgc atattgctga ggatgatggg gaggtggaca ccgatttccc cccgctggat
     1381 tccaatgagc ccattcataa gtttggcttc agtactttgg ccctggttga aaagtactca
     1441 tctcctggtc tgacatccaa agagtcactc tttgttcgaa taaatgctgc tcatggattc
     1501 tcccttattc aggtggacaa cacaaaggtt accatgaagg aaatcttact gaaggcagtg
     1561 aagcgaagaa aaggatccca gaaagtttca ggccctcagt accgcctgga gaagcagagc
     1621 gagcccaatg tcgccgttga cctggacagc actttggaga gccagagcgc atgggagttc
     1681 tgcctggtcc gcgagaacag ttcaagggca gacggggttt ttgaggagga ttcgcaaatt
     1741 gacatagcca cagtacagga tatgcttagc agccaccatt acaagtcatt caaagtcagc
     1801 atgatccaca gactgcgatt cacaaccgac gtacagctag gtatctctgg agacaaagta
     1861 gagatagacc ctgttacgaa tcagaaagcc agcactaagt tttggattaa gcagaaaccc
     1921 atctcaatcg attccgacct gctctgtgcc tgtgaccttg ctgaagagaa aagccccagt
     1981 cacgcaatat ttaaactcac gtatctaagc aatcacgact ataaacacct ctactttgaa
     2041 tcggacgctg ctaccgtcaa tgaaattgtg ctcaaggtta actacatcct ggaatcgcga
     2101 gctagcactg cccgggctga ctactttgct caaaaacaaa gaaaactgaa cagacgtacg
     2161 agcttcagct tccagaagga gaagaaatcc gggcagcagg taccgcgggc ccgggatcca
     2221 ccggtcgcca ccatgagcgg gggcgaggag ctgttcgccg gcatcgtgcc cgtgctgatc
     2281 gagctggacg gcgacgtgca cggccacaag ttcagcgtgc gcggcgaggg cgagggcgac
     2341 gccgactacg gcaagctgga gatcaagttc atctgcacca ccggcaagct gcccgtgccc
     2401 tggcccaccc tggtgaccac cctctgctac ggcatccagt gcttcgcccg ctaccccgag
     2461 cacatgaaga tgaacgactt cttcaagagc gccatgcccg agggctacat ccaggagcgc
     2521 accatccagt tccaggacga cggcaagtac aagacccgcg gcgaggtgaa gttcgagggc
     2581 gacaccctgg tgaaccgcat cgagctgaag ggcaaggact tcaaggagga cggcaacatc
     2641 ctgggccaca agctggagta cagcttcaac agccacaacg tgtacatccg ccccgacaag
     2701 gccaacaacg gcctggaggc taacttcaag acccgccaca acatcgaggg cggcggcgtg
     2761 cagctggccg accactacca gaccaacgtg cccctgggcg acggccccgt gctgatcccc
     2821 atcaaccact acctgagcac tcagaccaag atcagcaagg accgcaacga ggcccgcgac
     2881 cacatggtgc tcctggagtc cttcagcgcc tgctgccaca cccacggcat ggacgagctg
     2941 tacaggtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3001 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3061 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3121 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3181 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3241 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3301 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3361 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3421 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3481 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3541 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3601 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3841 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3901 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3961 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4021 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4081 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4141 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4681 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4981 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5221 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5281 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5341 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5401 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6241 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
