LOCUS       sequence-345751-        2954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of mouse tRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2954)
  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 2954)
  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: pBS-tRNA-Gly
FEATURES             Location/Qualifiers
     source          1..2954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(154..742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(234..253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(913..1704,1705..1773))
                     /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     1536..1555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1774..1878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1905..2360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1992..2011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2202..2223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2372..2710)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSKLTLTKGNKSWSSTAVAAAGAPPGNRTRVARMGILHD
                     TTTPAARYPIRPIVSRITIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPI
                     ALPNSCAA"
     primer_bind     2486..2508
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2500..2517
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2501..2517
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2524..2543
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2524..2542
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2667..2685)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2706..2722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2706..2722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2719..2741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2730..2746
                     /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)."
     promoter        complement(join(2754..2760,2761..2778,2779..2784))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2799..2820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2937..2954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
       61 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     1921 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1981 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2041 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2101 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2161 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2221 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2281 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2341 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2401 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2461 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2521 ttgtaatacg actcactata gggcgaattg ggtaccgcgc cgctggtgta gtggtatcat
     2581 gcaagattcc cattcttgcg acccgggttc gattcccggg cggcgcacca gcggccgcca
     2641 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     2701 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2761 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2941 ctcactgact cgct
//
