LOCUS       sequence-201171-        7296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged Sestrin2 in mammalian cells to make lentivirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7296)
  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 7296)
  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-Sestrin2
FEATURES             Location/Qualifiers
     source          1..7296
                     /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(3874..3894)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3879..4208
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4059..4194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4216..4614
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4783..5252
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5382..5399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5553..6141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5633..5652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6301..7092,7093..7161))
                     /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     6924..6943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7162..7266)
                     /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 tcgagatgat cgtggcggac tccgagtgcc
     2281 gcgcagagct caaggactac ctgcggttcg ccccgggcgg cgtcggcgac tcgggccccg
     2341 gagaggagca gagggagagc cgggctcggc gaggccctcg agggcccagc gccttcatcc
     2401 ccgtggagga ggtccttcgg gagggggctg agagcctcga gcagcacctg gggctggagg
     2461 cactgatgtc ctctgggcga gtagacaacc tggcagtggt gatgggcctg caccctgact
     2521 actttaccag cttctggcgc ctgcactacc tgctgctgca cacggatggt cccttggcca
     2581 gctcctggcg ccactacatt gccatcatgg ctgccgcccg ccatcagtgt tcttacctgg
     2641 taggctccca catggccgag tttctgcaga ctggtggtga ccctgagtgg ctgctgggcc
     2701 tccaccgggc ccccgagaag ctgcgcaaac tcagcgagat caacaagttg ctggcgcatc
     2761 ggccatggct catcaccaag gaacacatcc aggccttgct gaagaccggc gagcacactt
     2821 ggtccctggc cgagctcatt caggctctgg tcctgctcac ccactgccac tcgctctcct
     2881 ccttcgtgtt tggctgtggc atcctccctg agggggatgc agatggcagc cctgcccccc
     2941 aggcacctac accccctagt gaacagagca gccccccaag cagggacccg ttgaacaact
     3001 ctgggggctt tgagtctgcc cgcgacgtgg aggcgctgat ggagcgcatg cagcagctgc
     3061 aggagagcct gctgcgggat gaggggacgt cccaggagga gatggagagc cgctttgagc
     3121 tggagaagtc agagagcctg ctggtgaccc cctcagctga catcctggag ccctctccac
     3181 acccagacat gctgtgcttt gtggaagacc ctactttcgg atatgaggac ttcactcgga
     3241 gaggggctca ggcaccccct accttccggg cccaggatta tacctgggaa gaccatggct
     3301 actcgctgat ccagcggctt taccctgagg gtgggcagct gctggatgag aagttccagg
     3361 cagcctatag cctcacctac aataccatcg ccatgcacag tggtgtggac acctccgtgc
     3421 tccgcagggc catctggaac tatatccact gcgtctttgg catcagatat gatgactatg
     3481 attatgggga ggtgaaccag ctcctggagc ggaacctcaa ggtctatatc aagacagtgg
     3541 cctgctaccc agagaagacc acccgaagaa tgtacaacct cttctggagg cacttccgcc
     3601 actcagagaa ggtccacgtg aacttgctgc tcctggaggc gcgcatgcaa gccgctctgc
     3661 tgtacgccct ccgtgccatc acccgctaca tgacctgaga attctgcagt cgacggtacc
     3721 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     3781 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3841 aatgcaattc gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa
     3901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3961 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4141 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4201 tttgcaaaaa gcttcatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     4261 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     4321 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     4381 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     4441 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     4501 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     4561 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagatcga
     4621 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4681 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     4741 gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca
     4801 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg
     4861 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4921 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     4981 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     5041 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     5101 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     5161 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     5221 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     5281 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     5341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6241 tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7261 tccgcgcaca tttccccgaa aagtgccacc tgcagc
//
