LOCUS       sequence-197150-        7071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged SAMTOR in mammalian cells to make lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7071)
  AUTHORS   Gu X, Orozco JM, Saxton RA, Condon KJ, Liu GY, Krawczyk PA, Scaria 
            SM, Harper JW, Gygi SP, Sabatini DM
  TITLE     SAMTOR is an S-adenosylmethionine sensor for the mTORC1 pathway.
  JOURNAL   Science. 2017 Nov 10;358(6364):813-818. doi: 
            10.1126/science.aao3265.
  PUBMED    29123071
REFERENCE   2  (bases 1 to 7071)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-10- 10"; volume: "358"; 
            issue: "6364"; pages: "813-818"
COMMENT     Sequence Label: pLC242-GFP-SAMTOR
FEATURES             Location/Qualifiers
     source          1..7071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1386..1395
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1437..1458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1698..1717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2045..2066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2154..2174
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2187..2231
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3649..3669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3654..3983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3834..3969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3991..4389
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4558..5027
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5157..5174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5328..5916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5408..5427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6076..6867,6868..6936))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6699..6718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6937..7041)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccagtgtggt ggtacgctac
     1381 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2101 tgtacaagtc cggaggggat cggtggagct ccaccggtgg cggccgctct agagagaatc
     2161 tttattttca gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc
     2221 acatggacag cggaggtgga ggtactagtc tcgagatgga gccaggggcc ggcggccgaa
     2281 atactgctcg tgcgcagagg gccgggtccc cgaacactcc gccgccccgg gagcaggagc
     2341 ggaaactgga gcaggagaag ctctccggtg tggtgaagag cgtccaccgg cggctccgca
     2401 agaagtaccg agaagtggga gattttgata agatctggcg agaacactgt gaggatgaag
     2461 aaactctttg tgaatatgcc gttgcaatga aaaatttggc agataatcat tgggcaaaaa
     2521 cttgtgaggg cgaaggtcgt attgaatggt gttgtagtgt atgcagagaa tatttccaaa
     2581 atggtgggaa gagaaaagca cttgagaaag atgaaaaaag agctgtactt gccactaaga
     2641 ccactccagc cttaaatatg catgagtctt ctcaacttga aggtcattta accaacctca
     2701 gctttacaaa ccctgaattt ataactgagt tgctacaagc ctcaggaaaa atcagattac
     2761 ttgatgttgg cagctgcttt aacccatttc tgaagtttga agaatttcta actgttggca
     2821 tagatattgt acctgctgta gagagtgtct ataaatgtga tttcctgaac ttacagcttc
     2881 agcaaccact ccagcttgca caggatgcta tagatgcttt tttgaagcag ctgaaaaacc
     2941 ctattgattc tcttcctgga gagcttttcc atgtggttgt tttctctctc cttctttctt
     3001 attttccatc tccttaccag cgatggattt gctgcaagaa agcccatgaa ctgttagtgt
     3061 taaatggttt attactaatc atcacacctg attcctccca tcagaaccgt catgctatga
     3121 tgatgaaaag ctggaagatt gctatagagt ccctgggctt taaacgcttc aagtactcaa
     3181 aattttcaca tatgcatctg atggcattta ggaaaatctc tctaaaaacc acaagtgact
     3241 tggttagtag gaactaccca ggaatgttat atattcctca agatttcaac agtatagaag
     3301 atgaggaata ttctaaccct tcctgctatg ttcgatcaga tatagaagat gaacaactag
     3361 catatggttt cacagaactc cctgatgcgc catatgactc agattctgga gaaagtcaag
     3421 ccagctctat tcctttctat gagctagaag accccatatt acttttaagt taagaattct
     3481 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     3541 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3601 ctgaaacata aaatgaatgc aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc
     3661 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3901 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3961 ttggaggcct aggcttttgc aaaaagcttc atggccaagc ctttgtctca agaagaatcc
     4021 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     4081 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     4141 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     4201 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     4261 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     4321 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     4381 gagggctaag atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4441 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4501 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg
     4561 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4621 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4681 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     4741 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     4801 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     4861 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     4921 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     4981 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta
     5041 tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa
     5101 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5161 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5221 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5281 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5341 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5401 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5461 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5521 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5581 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5641 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5701 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5761 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5821 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5881 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5941 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6001 aaaaggatct tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta
     6061 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6121 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6181 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6241 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6301 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6361 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6421 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6481 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6541 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6601 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6661 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6721 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6781 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6841 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6961 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7021 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag c
//
