LOCUS       sequence-238751-        5875 bp    DNA     circular SYN 14-APR-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 Apr 14, 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        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"
     CDS             1825..2541
                     /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    2663..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2700..2719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2754..2773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2791..3246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2928..2949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3140..3159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3273..3377
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3379..3736
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3379..3570
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3402..3422)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3587..3722
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3771..4565
                     /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             3771..4565
                     /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(3825..3844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4435..4454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4753..4772)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4797..4844
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5173..5761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5662..5681
                     /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 accatgaagc agtcgatatt atctgtacgc
      661 ctagaacagt gccctctgca gctgaataac ccttttaacg agtattccaa atttgatggc
      721 aagggtcatg taggtacaac agcaaccaag aagatcgatg tctacctccc tctgcactcg
      781 agccaggaca gactgctgcc aatgaccgtg gtgacaatgg ccagcgccag ggtgcaggac
      841 ctgatcgggc tcatctgctg gcagtataca agcgaaggac gggagccgaa gctcaatgac
      901 aatgtcagtg cctactgcct gcatattgct gaggatgatg gggaggtgga caccgatttc
      961 cccccgctgg attccaatga gcccattcat aagtttggct tcagtacttt ggccctggtt
     1021 gaaaagtact catctcctgg tctgacatcc aaagagtcac tctttgttcg aataaatgct
     1081 gctcatggat tctcccttat tcaggtggac aacacaaagg ttaccatgaa ggaaatctta
     1141 ctgaaggcag tgaagcgaag aaaaggatcc cagaaagttt caggccctca gtaccgcctg
     1201 gagaagcaga gcgagcccaa tgtcgccgtt gacctggaca gcactttgga gagccagagc
     1261 gcatgggagt tctgcctggt ccgcgagaac agttcaaggg cagacggggt ttttgaggag
     1321 gattcgcaaa ttgacatagc cacagtacag gatatgctta gcagccacca ttacaagtca
     1381 ttcaaagtca gcatgatcca cagactgcga ttcacaaccg acgtacagct aggtatctct
     1441 ggagacaaag tagagataga ccctgttacg aatcagaaag ccagcactaa gttttggatt
     1501 aagcagaaac ccatctcaat cgattccgac ctgctctgtg cctgtgacct tgctgaagag
     1561 aaaagcccca gtcacgcaat atttaaactc acgtatctaa gcaatcacga ctataaacac
     1621 ctctactttg aatcggacgc tgctaccgtc aatgaaattg tgctcaaggt taactacatc
     1681 ctggaatcgc gagctagcac tgcccgggct gactactttg ctcaaaaaca aagaaaactg
     1741 aacagacgta cgagcttcag cttccagaag gagaagaaat ccgggcagca ggtaccgcgg
     1801 gcccgggatc caccggtcgc caccatgagc gggggcgagg agctgttcgc cggcatcgtg
     1861 cccgtgctga tcgagctgga cggcgacgtg cacggccaca agttcagcgt gcgcggcgag
     1921 ggcgagggcg acgccgacta cggcaagctg gagatcaagt tcatctgcac caccggcaag
     1981 ctgcccgtgc cctggcccac cctggtgacc accctctgct acggcatcca gtgcttcgcc
     2041 cgctaccccg agcacatgaa gatgaacgac ttcttcaaga gcgccatgcc cgagggctac
     2101 atccaggagc gcaccatcca gttccaggac gacggcaagt acaagacccg cggcgaggtg
     2161 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcaagga cttcaaggag
     2221 gacggcaaca tcctgggcca caagctggag tacagcttca acagccacaa cgtgtacatc
     2281 cgccccgaca aggccaacaa cggcctggag gctaacttca agacccgcca caacatcgag
     2341 ggcggcggcg tgcagctggc cgaccactac cagaccaacg tgcccctggg cgacggcccc
     2401 gtgctgatcc ccatcaacca ctacctgagc actcagacca agatcagcaa ggaccgcaac
     2461 gaggcccgcg accacatggt gctcctggag tccttcagcg cctgctgcca cacccacggc
     2521 atggacgagc tgtacaggta agcggccgcg actctagatc ataatcagcc ataccacatt
     2581 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2641 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2761 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2821 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2881 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2941 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3001 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3061 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3121 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3181 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3241 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3361 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3481 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3541 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3601 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3661 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3721 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3781 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3841 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3901 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3961 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4021 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4081 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4141 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4201 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4261 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4321 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4381 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4441 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4501 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4561 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4621 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4681 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4741 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4801 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4861 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4921 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4981 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5281 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
