LOCUS       sequence-200646-        6423 bp    DNA     circular SYN 15-APR-2025
DEFINITION  bacterial expression of mouse Sesn1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  AUTHORS   Lee JH, Budanov AV, Talukdar S, Park EJ, Park HL, Park HW, 
            Bandyopadhyay G, Li N, Aghajan M, Jang I, Wolfe AM, Perkins GA, 
            Ellisman MH, Bier E, Scadeng M, Foretz M, Viollet B, Olefsky J, 
            Karin M
  TITLE     Maintenance of metabolic homeostasis by Sestrin2 and Sestrin3.
  JOURNAL   Cell Metab. 2012 Sep 5;16(3):311-21. doi: 
            10.1016/j.cmet.2012.08.004.
  PUBMED    22958918
REFERENCE   2  (bases 1 to 6423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cmet.2012.08"; journalName: "Cell Metab"; date: 
            "2012-09-5- 5"; volume: "16"; issue: "3"; pages: "311-21"
COMMENT     Sequence Label: pGEX-4T-1 mSesn1 (1189)
FEATURES             Location/Qualifiers
     source          1..6423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        173..250
                     /gene="lacI"
                     /label=lacI promoter
     CDS             251..1333
                     /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(270..289)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1382..1387,1388..1405,1406..1412)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1420..1436
                     /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     1444..1460
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1444..1460
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1456..1983
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1476..1493)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1476..1492)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1485..1507)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2085..2090,2091..2106,2107..2113)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2121..2137
                     /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             2160..2813
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2771..2793
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2820..2837
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4375..4397)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4542..4560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4628..4732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4733..4801,4802..5593)
                     /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(4951..4970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5764..6352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6253..6272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
       61 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      121 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
      181 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
      241 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      301 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
      361 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
      421 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
      481 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
      541 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
      601 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
      661 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
      721 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
      781 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
      841 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
      901 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
      961 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1021 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1081 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1141 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1201 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     1261 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1321 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     1381 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     1441 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     1501 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1561 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1621 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     1681 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     1741 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     1801 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     1861 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     1921 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     1981 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     2041 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     2101 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
     2161 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     2221 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     2281 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     2341 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     2401 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     2461 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     2521 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     2581 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     2641 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     2701 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     2761 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     2821 tggttccgcg tggatccatg cgcctggccg ccgcgtcgaa cgaggcgtac gcagcctcgc
     2881 tggctgtctc cgagctgctg agctgccacc agtgcggcgg ggaccgcggc caggacgagg
     2941 aacttggaat cagaatccct cggccactag gacacggacc aagcaggttc atcccagaga
     3001 aggagatgct gcaggtgggc agtgaagacg cacagatgca tgcgttattt gcagactcct
     3061 ttgctgcttt gggtcgtctg gataacatca cattagtgat ggttttccat ccacaatatt
     3121 tagaaagttt cttaaagact caacactacc tactacaaat ggatgggcca ttacccctcc
     3181 attatcggca ctacattgga atcatggcgg cagcgaggca tcagtgttcc tacctggtga
     3241 atctgcatgt cagtgacttt cttcatgttg gtggggaccc caagtggctc aatggtttag
     3301 agaatgctcc tcaaaaactg cagaacttag gagaacttaa caaagtgtta gcccataggc
     3361 cttggctgat taccaaagaa catatcgagg gccttctgaa agcagaggag cacagctggt
     3421 ctctcgccga gctggtccat gccgtggttc tgctcacaca ctaccactct cttgcctcct
     3481 tcacatttgg ctgtggaatt agtccagaga ttcattgcga tggtggccac accttcagac
     3541 cgccttctgt tagtaactac tgcatctgtg acattacaaa tggcaatcac agtgtggatg
     3601 agatgcaagt caactcagca ggaaatgctt cggtaagtga ttccttcttt gaggtggaag
     3661 ctctcatgga aaagatgagg cagttacagg aatgccgaga ggaggaggag gctagccagg
     3721 aggagatggc gtcgcggttt gagatggaaa agagagagag catgtttgtc ttctcttcgg
     3781 acgatgatga agttacacca gcaagagatg tgtctcggca ctttgaggac actagttacg
     3841 gctacaagga tttctccaga cacgggatgc atgtcccaac gtttcgtgtc caggactact
     3901 gctgggaaga ccatggttac tctctggtga atcgtctcta tccagatgtg ggacagttaa
     3961 ttgatgagaa atttcacatt ccttacaatc taacttacaa tactatggcc atgcacaaag
     4021 atgttgatac ctcaatgctc agacgggcta tctggaacta tatccactgc atgtttggaa
     4081 tacgatacga cgactatgac tatggggaaa ttaaccagct gctggatcgt agctttaaag
     4141 tttacatcaa aaccgtcgtg tgcactcctg aaaaggttac caaaaggatg tatgacagct
     4201 tctggaggca gttcaagcac tctgagaagg tccatgttaa tctgcttctt atagaagcta
     4261 ggatgcaagc agaactcctg tatgctctga gagccattac ccggtatatg acctgagaat
     4321 tcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     4381 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4441 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     4501 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     4561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     5281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6421 tgc
//
