LOCUS       sequence-493032-        3079 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Plasmid for in vitro expression of M1 RNA of E. coli RNase P.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3079)
  AUTHORS   Miyachi R, Masuda K, Shimizu Y, Ichihashi N
  TITLE     Simultaneous in vitro expression of minimal 21 transfer RNAs by tRNA
            array method.
  JOURNAL   Nat Commun. 2025 Aug 26;16(1):7418. doi: 10.1038/s41467-025-62588-y.
  PUBMED    40858540
REFERENCE   2  (bases 1 to 3079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-08-26"; volume: "16(1):7418. doi"; pages: " 
            10.1038/s41467-025-62588-y"
COMMENT     Sequence Label: pUC-M1
FEATURES             Location/Qualifiers
     source          1..3079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      785..1373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1274..1293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1527..1544
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1661..1682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1697..1702,1703..1720,1721..1727)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1735..1751
                     /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     1740..1762
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1759..1775
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1759..1775
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2238..2255)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2238..2254)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2247..2269)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2463..2482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2582..2604
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2642..2660)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2728..2832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2833..2901,2902..3079,1..614)
                     /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     complement(3051..3070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
       61 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      121 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      181 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      241 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      301 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      361 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      421 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      481 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      541 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      601 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      661 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      721 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      781 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      841 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      901 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      961 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1021 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1081 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1141 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1201 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1261 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1321 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1381 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1441 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1501 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1561 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1621 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     1681 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     1741 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc
     1801 tgcaggtcga ctctagagga tccggatgac gtaaatcagg tgaaactgac cgataagccg
     1861 ggttctgtcg tggacagtca ttcatctagg ccagcaatcg ctcactggct caagcagcct
     1921 acccgggttc agtacgggcc gtaccttatg aacccctatt tggccttgct ccgggtggag
     1981 tttaccgtgc cacggactgt taccagccgc gcggtgcgct cttaccgcac cctttcaccc
     2041 ttacctgatc ccgcttgcgc gggccatcgg cggtttgctc tctgttgcac tggtcgtggg
     2101 tttccccccc aggcgttacc tggcaccctg ccctatggag cccggacttt cctcccctcc
     2161 gcccgtctcc cccgaagagg acgacgacga agcggcgact gtctggtcag cttctatagt
     2221 gagtcgtatt agaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2281 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2341 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     2401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     2461 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2521 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2581 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2641 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2701 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3061 gcggtattat cccgtattg
//
