LOCUS       sequence-288137-        6493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of SNIPR RNA with a LacZ reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  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 6493)
  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_sensor_LacZ
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             148..3222
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(168..185)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(168..184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(177..199)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3223..3269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3499..4314)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3499..4314)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4222..4241
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4315..4406)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4424..5059)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(4569..4588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5257..6339)
                     /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     6301..6320
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6340..6417)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6477..6493,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6477..6493,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggtagaatg aaatgaataa gacaataaca gtaacatggt aggtaagtgt gataacatca
       61 tagaggagat acagaatgtt atcacactta cctaccgaac aagaactccg acaaatcatg
      121 cgtaaaggag aagaactttt cactggaatg accatgatta cggattcact ggccgtcgtt
      181 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      241 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      301 ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg
      361 gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg
      421 cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat
      481 ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat
      541 gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttaa ctcggcgttt
      601 catctgtggt gcaacgggcg ctgggtcggt tacggccagg acagtcgttt gccgtctgaa
      661 tttgacctga gcgcattttt acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgc
      721 tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg cattttccgt
      781 gacgtctcgt tgctgcataa accgactaca caaatcagcg atttccatgt tgccactcgc
      841 tttaatgatg atttcagccg cgctgtactg gaggctgaag ttcagatgtg cggcgagttg
      901 cgtgactacc tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc
      961 accgcgcctt tcggcggtga aattatcgat gagcgtggtg gttatgccga tcgcgtcaca
     1021 ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt
     1081 gcggtggttg aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc
     1141 ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg
     1201 attcgaggcg ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag
     1261 acgatggtgc aggatatcct gctgatgaag cagaacaact ttaacgccgt gcgctgttcg
     1321 cattatccga accatccgct gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg
     1381 gatgaagcca atattgaaac ccacggcatg gtgccaatga atcgtctgac cgatgatccg
     1441 cgctggctac cggcgatgag cgaacgcgta acgcgaatgg tgcagcgcga tcgtaatcac
     1501 ccgagtgtga tcatctggtc gctggggaat gaatcaggcc acggcgctaa tcacgacgcg
     1561 ctgtatcgct ggatcaaatc tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga
     1621 gccgacacca cggccaccga tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag
     1681 cccttcccgg ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc tggagagacg
     1741 cgcccgctga tcctttgcga atacgcccac gcgatgggta acagtcttgg cggtttcgct
     1801 aaatactggc aggcgtttcg tcagtatccc cgtttacagg gcggcttcgt ctgggactgg
     1861 gtggatcagt cgctgattaa atatgatgaa aacggcaacc cgtggtcggc ttacggcggt
     1921 gattttggcg atacgccgaa cgatcgccag ttctgtatga acggtctggt ctttgccgac
     1981 cgcacgccgc atccagcgct gacggaagca aaacaccagc agcagttttt ccagttccgt
     2041 ttatccgggc aaaccatcga agtgaccagc gaatacctgt tccgtcatag cgataacgag
     2101 ctcctgcact ggatggtggc gctggatggt aagccgctgg caagcggtga agtgcctctg
     2161 gatgtcgctc cacaaggtaa acagttgatt gaactgcctg aactaccgca gccggagagc
     2221 gccgggcaac tctggctcac agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa
     2281 gccgggcaca tcagcgcctg gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc
     2341 cccgccgcgt cccacgccat cccgcatctg accaccagcg aaatggattt ttgcatcgag
     2401 ctgggtaata agcgttggca atttaaccgc cagtcaggct ttctttcaca gatgtggatt
     2461 ggcgataaaa aacaactgct gacgccgctg cgcgatcagt tcacccgtgc accgctggat
     2521 aacgacattg gcgtaagtga agcgacccgc attgacccta acgcctgggt cgaacgctgg
     2581 aaggcggcgg gccattacca ggccgaagca gcgttgttgc agtgcacggc agatacactt
     2641 gctgatgcgg tgctgattac gaccgctcac gcgtggcagc atcaggggaa aaccttattt
     2701 atcagccgga aaacctaccg gattgatggt agtggtcaaa tggcgattac cgttgatgtt
     2761 gaagtggcga gcgatacacc gcatccggcg cggattggcc tgaactgcca gctggcgcag
     2821 gtagcagagc gggtaaactg gctcggatta gggccgcaag aaaactatcc cgaccgcctt
     2881 actgccgcct gttttgaccg ctgggatctg ccattgtcag acatgtatac cccgtacgtc
     2941 ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat tgaattatgg cccacaccag
     3001 tggcgcggcg acttccagtt caacatcagc cgctacagtc aacagcaact gatggaaacc
     3061 agccatcgcc atctgctgca cgcggaagaa ggcacatggc tgaatatcga cggtttccat
     3121 atggggattg gtggcgacga ctcctggagc ccgtcagtat cggcggaatt ccagctgagc
     3181 gccggtcgct accattacca gttggtctgg tgtcaaaaat aatagcataa ccccttgggg
     3241 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
     3301 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     3361 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
     3421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3481 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3541 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3601 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3661 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3721 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     3781 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3841 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
     3901 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3961 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     4021 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4081 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4141 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     4201 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4261 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     4321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4381 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg
     4441 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct
     4501 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     4561 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc
     4621 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt
     4681 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg
     4741 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact
     4801 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt
     4861 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag
     4921 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg
     4981 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct
     5041 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc
     5101 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     5161 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5221 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5281 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5341 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5401 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5461 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5521 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5581 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5641 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5701 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5761 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5821 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5881 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5941 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6001 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6061 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6121 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6181 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6241 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6301 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6361 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6421 gatctcgacg ctctccctta tggccgggtt tagaaatctg aagctctaga gagcgctaat
     6481 acgactcact ata
//
