LOCUS       sequence-200648-        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of dSesn .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  AUTHORS   Lee JH, Budanov AV, Park EJ, Birse R, Kim TE, Perkins GA, Ocorr K, 
            Ellisman MH, Bodmer R, Bier E, Karin M
  TITLE     Sestrin as a feedback inhibitor of TOR that prevents age-related 
            pathologies.
  JOURNAL   Science. 2010 Mar 5;327(5970):1223-8. doi: 10.1126/science.1182228.
  PUBMED    20203043
REFERENCE   2  (bases 1 to 6454)
  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: "2010-03-5- 5"; volume: "327"; issue: 
            "5970"; pages: "1223-8"
COMMENT     Sequence Label: pGEX-4T-1 dSesn (1187)
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..71
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        144..221
                     /gene="lacI"
                     /label=lacI promoter
     CDS             222..1304
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(241..260)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1353..1358,1359..1376,1377..1383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1391..1407
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1415..1431
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1415..1431
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1427..1954
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1447..1464)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1447..1463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1456..1478)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2056..2061,2062..2077,2078..2084)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2092..2108
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             2131..2784
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2742..2764
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2791..2808
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4377..4399)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4544..4562)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4630..4734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4735..4803,4804..5595)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4953..4972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5766..6354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6255..6274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
       61 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
      121 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
      181 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
      241 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
      301 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
      361 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
      421 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
      481 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
      541 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
      601 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
      661 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
      721 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
      781 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
      841 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
      901 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
      961 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     1021 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     1081 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     1141 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     1201 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     1261 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     1321 gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     1381 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     1441 ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1501 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1561 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     1621 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
     1681 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
     1741 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
     1801 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
     1861 tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
     1921 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
     1981 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
     2041 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg
     2101 gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta ttggaaaatt
     2161 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
     2221 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
     2281 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
     2341 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
     2401 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
     2461 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
     2521 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
     2581 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
     2641 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
     2701 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
     2761 ggtggtggcg accatcctcc aaaatcggat ctggttccgc gtggatcccc ggaattcatg
     2821 tactacgccg tcgattacta cgcggatatg ggacaaatct ctcaagattg tttcgctgca
     2881 acgcaaactg gagcatccga ctttgacatg gacgagctcg atgacctgga ccaggtgacc
     2941 caagtgattg gatatcatcc ccagttccac gatcacttcc tggccaccca gaacttcatt
     3001 atgaagggcg atggtccgct gcctaacgat tacagatact acttggccat aattgctgct
     3061 gcccgccatc agtgtccgta tctggtgaag cgctacgaga aggagttcat caaccagggc
     3121 ggcgatagtg cctggctggg cggcctggac tttatacccg ccaaactacg tgccatatac
     3181 gatatcaata aaatattagc ccatcggccc tggctgctgc gcaaggagca tatcgagcgg
     3241 ctcaccaagg ggaagaacag ctggtcgctg tcggaggtgg tgcacgccat ggtcctgctc
     3301 tcgcacttcc actcgctgtc ctcctttgtg ttctcctgcg gcctcacaca aaagctggac
     3361 gggctgtcca gtcccaagct gaagagtccg cccgcagcag tggcggccct cgcacccacc
     3421 attctcatca ctcccacctc gccgacggag ccgcagaagg ggaagccagt gctggcggag
     3481 atctcgctca acaatgcaaa tccggattac gacagccaga ccgcagcgag cagtaatgga
     3541 ggagctccac cggactcggc aaatgcggtt gcggatggac ccgatgcaac gacccttaac
     3601 ggatatttgg ccacggcgca gcagctgccg cagcagcacg gcatcagtgt ggagacgcta
     3661 atggagcgca tgaaggtcct gtcccagaag caggacgagt gcagcgaggc ggagctgagc
     3721 agccgcttcc agaaggtgga gcagcagacg gcggagctgg cggccgtgac tccggaggcg
     3781 gctgtgggcg tgcccaccaa cctgtcgcac tacgtggacg atgccaactt catttaccag
     3841 gactttgccc ggcgcggcac cgagagcatc aacacgtttc gcatccagga ctactcttgg
     3901 gaggatcacg gctactcgct agtggacgga ttgtacaacg atgtgggcat cttcttggac
     3961 gccaagtttc gggcggccta caatctcacc tactgcacca tgggcggcat caagaatgtg
     4021 gacacgtcca agttccggcg ggccatttgg aactacatcc agtgcatcta cggcattcgc
     4081 cacgacgact acgactatgg cgaagtgaac cagctcctgg tgcgtcccct gaaaatgttt
     4141 attaagacag cctgctgctt ccccgagcga atcaccacca aggactacga tagtgtgctg
     4201 gtcgagctgc aggacagtga aaaggttcat gtgaatctga tgataatgga agcccgtaat
     4261 caggccgagt tactctatgc tctgcgcgag ataatgcgct atatgacttg actcgagcgg
     4321 ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4381 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4441 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4501 tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta
     4561 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     4981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6421 cctgcgttat cccctgattc tgtggataac cgta
//
