LOCUS       sequence-200645-        6387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of mouse Sesn2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6387)
  AUTHORS   Park HW, Park H, Ro SH, Jang I, Semple IA, Kim DN, Kim M, Nam M, 
            Zhang D, Yin L, Lee JH
  TITLE     Hepatoprotective role of Sestrin2 against chronic ER stress.
  JOURNAL   Nat Commun. 2014 Jun 20;5:4233. doi: 10.1038/ncomms5233.
  PUBMED    24947615
REFERENCE   2  (bases 1 to 6387)
  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"; journalName: "Nat 
            Commun."; date: "2014-06-20"; volume: "5:4233. doi"; pages: " 
            10.1038/ncomms5233"
COMMENT     Sequence Label: pGEX-4T-1 mSesn2 (1190)
FEATURES             Location/Qualifiers
     source          1..6387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        207..284
                     /gene="lacI"
                     /label=lacI promoter
     CDS             285..1367
                     /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(304..323)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1416..1421,1422..1439,1440..1446)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1454..1470
                     /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     1478..1494
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1478..1494
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1490..2017
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1510..1527)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1510..1526)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1519..1541)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2119..2124,2125..2140,2141..2147)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2155..2171
                     /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             2194..2847
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2805..2827
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2854..2871
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4373..4395)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4540..4558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4626..4730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4731..4799,4800..5591)
                     /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(4949..4968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5762..6350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6251..6270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      121 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      181 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
      241 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
      301 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
      361 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
      421 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
      481 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
      541 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
      601 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
      661 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
      721 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
      781 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
      841 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
      901 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
      961 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     1021 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     1081 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     1141 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     1201 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     1261 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1321 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1381 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     1441 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     1501 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     1561 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     1621 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     1681 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     1741 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     1801 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     1861 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     1921 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1981 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     2041 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     2101 atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
     2161 gcggataaca atttcacaca ggaaacagta ttcatgtccc ctatactagg ttattggaaa
     2221 attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa
     2281 gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt
     2341 ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg
     2401 gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt
     2461 gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt
     2521 gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg
     2581 ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc
     2641 catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc
     2701 ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt
     2761 gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg
     2821 tttggtggtg gcgaccatcc tccaaaatcg gatctggttc cgcgtggatc catgatcgta
     2881 gcggactccg agtgccattc cgagatcaag ggctacctgc cgttcaccag gggtggtgtg
     2941 gccggccccg agacccgaga ggagcatcgg gagggccagg ctcggcgagg ctctcgaggg
     3001 cccagcgctt tcattccagt ggaagagatc cttcgggaag gagctgagag cctggagcag
     3061 cacctggggc tcgaggccct gatgtcctca gggcgggtgg acaacctggc ggtggtgatg
     3121 ggtctacacc cggactacct tagcagcttc tggcgcctgc attacctgct gctgcatacg
     3181 gatgggcccc tggccagctc ctggcgccac tacattgcca tcatggctgc tgcccgtcac
     3241 cagtgttctt acctggtggg ttcccacatg actgagttcc tgcagaccgg cggtgacccc
     3301 gagtggctgc tgggccttca ccgggcccct gagaagctcc gcaagctcag cgaggtcaac
     3361 aagttactgg cgcatcggcc gtggctcatc accaaggagc atatccaggc cttgctgaag
     3421 actggcgagc acagctggtc cctggccgaa ctcatccaag cactggtcct gcttacccac
     3481 tgccactctc tggcctcctt tgtgtttggc tgtggaatac ttcctgaggg agatgcagag
     3541 ggcagccctg cctcccaggc accttcgcct cccagtgagc aaggcacccc cccgagcggg
     3601 gacccactga acaactcagg gggctttgaa gccgctcgtg acgtagaagc tctgatggaa
     3661 cgcatgaggc agcttcagga gagcctgctg cgggatgagg gggcttcgca ggaggaaatg
     3721 gagaaccgtt ttgagctgga gaagtcagaa agcctgctgg tgaccccctc agcggatatc
     3781 ctggagcctt ctccacaccc agacatccta tgctttgtgg aggaccccgc atttggatat
     3841 gaagacttca ctcgtcgagg gactcaggcg cccccaacct tccgtgccca ggattatacc
     3901 tgggaagacc atggctactc gctgatccag aggctctacc ctgagggagg gcagctgctg
     3961 gatgagaagt tccaggtagc ctgcagcctc acctataaca ccatcgccat gcacagtggc
     4021 gtggacacct ccatgctccg cagggccatc tggaactaca tccactgcgt ctttggcatc
     4081 agatacgatg actatgatta cggcgaggta aaccagctcc tggaacggaa cctcaaaatc
     4141 tatatcaaga cggtggcctg ctaccctgag aagacgaccc gtaggatgta caacctcttt
     4201 tggaggcact tccgccactc agagaaggtt catgtgaact tgctgctcct tgaagcccgc
     4261 atgcaagcgg ccctgctcta tgccctgcgc gccatcaccc ggtacatgac ctgagaattc
     4321 atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4381 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4441 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     4501 gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     4561 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4681 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4741 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4801 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4861 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4921 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     4981 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5041 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5101 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5161 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5221 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     5281 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5341 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5401 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5461 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5521 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5581 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5641 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5701 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5761 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5821 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5881 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5941 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6001 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6061 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6121 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6181 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6241 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6301 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6361 acgcggcctt tttacggttc ctggcct
//
