LOCUS       sequence-275326-        5603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of a non-cognate SNIPR trigger RNA with a 
            mutation at position 36.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5603)
  AUTHORS   Hong F, Ma D, Wu K, Mina LA, Luiten RC, Liu Y, Yan H, Green AA
  TITLE     Precise and Programmable Detection of Mutations Using Ultraspecific 
            Riboregulators.
  JOURNAL   Cell. 2020 Feb 25. pii: S0092-8674(20)30155-0. doi: 
            10.1016/j.cell.2020.02.011.
  PUBMED    32109416
REFERENCE   2  (bases 1 to 5603)
  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: "Cell. 2020 
            Feb 25. pii: S0092-8674(20)30155-0. doi: 
            10.1016/j.cell.2020.02.011."
COMMENT     Sequence Label: p_SNIPR_target_mut_36
FEATURES             Location/Qualifiers
     source          1..5603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      80..126
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(301..306,307..323,324..329))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(356..374)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        442..546
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(547..615,616..1407)
                     /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(765..784)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1578..2166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2067..2086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2320..2337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2352..2491
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2417..2436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2577..2599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2593..2784)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4096..5178)
                     /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     5140..5159
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5179..5256)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5462..5481
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5587..5603,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5587..5603,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggtgaatat cctattcgat attcacaata tcacatttac ctaccatgtc aatcacttct
       61 tattcatttc attctatctt agcataaccc cttggggcct ctaaacgggt cttgaggggt
      121 tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca
      181 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
      241 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
      301 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
      361 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      421 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      481 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      541 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      601 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      661 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      721 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      781 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      841 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      901 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      961 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1021 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1081 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1141 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1201 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1261 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1321 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1381 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1441 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1501 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1561 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1621 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1681 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1741 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1801 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1861 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1921 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1981 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2041 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2101 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2161 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2221 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2281 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2341 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2401 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     2461 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     2521 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     2581 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     2641 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     2701 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     2761 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     2821 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     2881 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     2941 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     3001 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     3061 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     3121 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     3181 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     3241 gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc tgctggagat
     3301 ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa
     3361 gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc
     3421 attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg
     3481 tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa
     3541 atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat
     3601 ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg
     3661 ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc
     3721 gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc gataatggcc
     3781 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     3841 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     3901 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     3961 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4021 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4081 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4141 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     4201 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     4261 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     4321 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     4381 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     4441 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     4501 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     4561 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     4621 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     4681 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     4741 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     4801 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4861 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4921 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     4981 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5041 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5101 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     5161 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     5221 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     5281 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     5341 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     5401 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     5461 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     5521 cgatgcgtcc ggcgtagagg atcgagatct cggccgggtt tagaaatctg aagctctaga
     5581 gagcgctaat acgactcact ata
//
