LOCUS       sequence-288138-        4211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of SNIPR RNA with a GFP-ASV reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4211)
  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 4211)
  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_GFP
FEATURES             Location/Qualifiers
     source          1..4211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             118..831
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             118..831
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(148..176)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     771..791
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(926..944)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      940..987
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1217..2032)
                     /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(1217..2032)
                     /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     1940..1959
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2033..2124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2142..2777)
                     /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(2287..2306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2975..4057)
                     /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     4019..4038
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4058..4135)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4195..4211,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4195..4211,1..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 cactggagtt gtcccaattc ttgttgaatt agatggtgat
      181 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
      241 cttaccctta aatttatttg cactactgga aaactacctg ttccgtggcc aacacttgtc
      301 actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcacat gaaacagcat
      361 gactttttca agagtgccat gcccgaaggt tacgtacagg aaagaactat atttttcaaa
      421 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
      481 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
      541 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
      601 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
      661 tatcaacaaa atactccgat tggcgatggc cctgtccttt taccagacaa ccattacctg
      721 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
      781 gagtttgtaa ccgctgctgg gattacacat ggcatggatg aactatacaa aaggcctgca
      841 gcaaacgacg aaaactacgc tgcatcagtt taataagata aaccagagcg gcacggcaag
      901 cagagtatac gagattcggt agccaccgct gagcaataac tagcataacc ccttggggcc
      961 tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc
     1021 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     1081 cgaccctgcc ctgaaccgac gacaagctga cgaccgggtc tccgcaagtg gcacttttcg
     1141 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1201 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1261 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1321 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1381 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1441 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     1501 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1561 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg gtcgctgtta aaaggacaat
     1621 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1681 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     1741 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1801 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1861 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     1921 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     1981 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc atactcttcc
     2041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2101 aatgtattta gaaaaataaa caaataggca tgctagcgca gaaacgtcct agaagatgcc
     2161 aggaggatac ttagcagaga gacaataagg ccggagcgaa gccgtttttc cataggctcc
     2221 gcccccctga cgaacatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     2281 gactataaag ataccaggcg tttccccctg atggctccct cttgcgctct cctgttcccg
     2341 tcctgcggcg tccgtgttgt ggtggaggct ttacccaaat caccacgtcc cgttccgtgt
     2401 agacagttcg ctccaagctg ggctgtgtgc aagaaccccc cgttcagccc gactgctgcg
     2461 ccttatccgg taactatcat cttgagtcca acccggaaag acacgacaaa acgccactgg
     2521 cagcagccat tggtaactga gaattagtgg atttagatat cgagagtctt gaagtggtgg
     2581 cctaacagag gctacactga aaggacagta tttggtatct gcgctccact aaagccagtt
     2641 accaggttaa gcagttcccc aactgactta accttcgatc aaaccgcctc cccaggcggt
     2701 tttttcgttt acagagcagg agattacgac gatcgtaaaa ggatctcaag aagatccttt
     2761 acggattccc gacaccatca ctctagattt cagtgcaatt tatctcttca aatgtagcac
     2821 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     2881 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     2941 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3061 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3121 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     3181 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     3241 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     3301 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     3361 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     3421 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     3481 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3541 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3601 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3661 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3721 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3781 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3841 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3901 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3961 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4021 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4081 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     4141 tctcgacgct ctcccttatg gccgggttta gaaatctgaa gctctagaga gcgctaatac
     4201 gactcactat a
//
