LOCUS       sequence-345153-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of MBP-tagged mouse SLFN2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  AUTHORS   Yue T, Zhan X, Zhang D, Jain R, Wang KW, Choi JH, Misawa T, Su L, 
            Quan J, Hildebrand S, Xu D, Li X, Turer E, Sun L, Moresco EMY, 
            Beutler B
  TITLE     SLFN2 protection of tRNAs from stress-induced cleavage is essential 
            for T cell-mediated immunity.
  JOURNAL   Science. 2021 May 14;372(6543). pii: 372/6543/eaba4220. doi: 
            10.1126/science.aba4220.
  PUBMED    33986151
REFERENCE   2  (bases 1 to 7759)
  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: "Science."; 
            date: "2021-05-14"; volume: "372(6543). pii: 372/6543/eaba4220. 
            doi"; pages: " 10.1126/science.aba4220"
COMMENT     Sequence Label: pMal-SLFN2
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(329..334,335..350,351..357)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    365..381
                     /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     370..392
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             451..1551
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1525..1548
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1600..1614
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(2773..2790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2773..2789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2782..2804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2968..2985)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3138..3224
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3316..3343
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3362..3453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3454..3522,3523..4314)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3672..3691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4356..4869)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4551..4572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4763..4782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4980..5568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5469..5488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5722..5739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5754..5896
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5822..5841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5982..6004
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5998..6189)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6685..6762
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(6763..7759,1..86)
                     /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(6782..6801)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
       61 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
      121 taggcacaat tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg
      181 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat
      241 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg
      301 gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt
      361 ggaattgtga gcggataaca atttcacaca ggaaacagcc agtccgttta ggtgttttca
      421 cgagcacttc accaacaagg accatagcat atgaaaactg aagaaggtaa actggtaatc
      481 tggattaacg gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa
      541 gataccggaa ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag
      601 gttgcggcaa ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc
      661 tacgctcaat ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg
      721 tatccgttta cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct
      781 gttgaagcgt tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg
      841 gaagagatcc cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc
      901 aacctgcaag aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc
      961 aagtatgaaa acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa
     1021 gcgggtctga ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat
     1081 tactccatcg cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg
     1141 tgggcatggt ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc
     1201 ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc
     1261 agtccgaaca aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt
     1321 ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa
     1381 gagttggcga aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc
     1441 atgccgaaca tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac
     1501 gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc
     1561 tcgaacaaca acaacaataa caataacaac aacctcgggg atgacgatga caaggtaccg
     1621 gaattcatgg gtactagact tgaggcaact gagcaaagca accacagaac tcagaggaat
     1681 gacattcagc tggaaaatgc aaaggctgct gggaaaatgg gcatcagtgt tgatctggaa
     1741 gccaaatatg ctaagctggg tctaaatctc ggagcaatca cttttggaga gaaggatagg
     1801 aagaaaatga agaattctca cctcagaaaa caggagaatg caaacatctc tctagctgta
     1861 tgtgctctcc tgaattcggg aggtggagca atcaaggtta aaattgaaaa tgaaaattat
     1921 agtctcacta gggatggcct gggactagat ttggaagcct ctctttgtaa atgtctgccc
     1981 tttgtccagt ggcacctgga cttcacggag agcgaaggct acatttatat ctacgtgaaa
     2041 tcgtggagcc aagaaatctt tgggctgcct attggcaccc taagaaccaa tttgtatgta
     2101 aggagcatgt catcttctgt acaagtgagc gccgctgctg ccctggaatt tctccaggac
     2161 ctggaggaaa ctggagggag accctgtgtc agaccagagt tgcctgcaag catagccttc
     2221 cctgaagtgg aaggagaatg gcacctggag gatttggctg ctgcattgtt taacaggaca
     2281 gaatttcagt acgaggaaac tttccccttt accagatcca gatatgttga agttacattg
     2341 ctttcagcga aacgcctgcg aaaacgcatc aaagagctcc tccctcaaac tgtttctgct
     2401 tttgcaaaca cggatggggg atttttgttc attggtttgg atggcaaaac ccagcaaatt
     2461 attggttttg aagcagagaa gagcgatctc gtgcttctag agagtgaaat agaaaagcac
     2521 atccggcagc tgcctgtcac tcacttctgt gaggagaagg agaagatcaa gtacacgtgc
     2581 aaattcatcg aagtgcacaa atccggagct gtgtgtgcat atgtgtgtgc gctcagagtg
     2641 gagagattct gctgtgcagt attcgctgca gagcccgagt cctggcatgt ggaaggcggc
     2701 tgtgtgaaga ggtttaccac agaggaatgg gtgaaactcc agatgaatgc cccatcaggt
     2761 tgaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2821 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2881 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc agcttggctg
     2941 ttttggcgga tgagataaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     3001 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     3061 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     3121 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     3181 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     3241 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     3301 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     3361 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3421 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3481 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3541 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3601 aacagcggta agatccttga gagttttcgc cccgaagaac gtttcccaat gatgagcact
     3661 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     3721 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3781 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3841 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3901 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3961 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4021 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4081 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4141 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4201 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4261 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4321 gaccaagttt actcatatat actttagatt gatttacccc ggttgataat cagaaaagcc
     4381 ccaaaaacag gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa
     4441 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4501 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4561 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4621 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     4681 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4741 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4801 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4861 agggcgcgta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4921 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4981 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5041 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5101 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5161 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5221 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5281 gcgcagcggt cgggctgaac ggggggttcg tgcacagagc ccagcttgga gcgaacgacc
     5341 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5401 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5461 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5521 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5581 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5641 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5701 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5761 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca
     5821 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     5881 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5941 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6001 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     6061 gtcgtgcagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     6121 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     6181 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     6241 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     6301 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     6361 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     6421 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     6481 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     6541 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     6601 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     6661 ggacccaacg ctgcccgaaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg
     6721 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     6781 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     6841 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     6901 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     6961 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     7021 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     7081 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     7141 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     7201 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     7261 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     7321 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     7381 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     7441 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     7501 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     7561 gatatctcag tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     7621 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     7681 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     7741 aaaaccaccc tggcgccca
//
