LOCUS       sequence-445452-        4501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in response to D-Ribose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4501)
  AUTHORS   d'Oelsnitz S, Love JD, Ellington AD, Ross D
  TITLE     Ligify: Automated Genome Mining for Ligand-Inducible Transcription 
            Factors.
  JOURNAL   ACS Synth Biol. 2024 Aug 16;13(8):2577-2586. doi: 
            10.1021/acssynbio.4c00372. Epub 2024 Jul 19.
  PUBMED    39029917
REFERENCE   2  (bases 1 to 4501)
  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"; doi: 
            "10.1021/acssynbio.4c00372"; journalName: "ACS Synth Biol"; date: 
            "2024-08-16- 16"; volume: "13"; issue: "8"; pages: "2577-2586"
COMMENT     Sequence Label: pLigify_dRibose
FEATURES             Location/Qualifiers
     source          1..4501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        418..467
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(1714..1730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1714..1730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1727..1749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1910..2455)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1990..2009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2566..3381)
                     /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(2566..3381)
                     /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     3289..3308
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3373..3392
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3502..3518
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3502..3518
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3519..3565
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3679..3701
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3693..3710
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3694..3710
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(3845..4501,1..57))
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3885..3905)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     join(4500..4501,1..27)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt tactcatata
       61 taccccctta ttctcccgta ttaaacaaaa ttatttgtag aggccccatt tcgtcctttt
      121 ggactcatca ggggtggtac acaccaccct atggggcttg gcttcctcct ttcaattgtg
      181 ggaacgttct cacatttaca tgtatgattg tatccgtttt catttatttt gtcaatcctt
      241 tttatatttc gttttaataa caaaaaagcc tacttttcag taggcctttt aaattttctt
      301 gcaaaaaaaa accccgccga agcggggttt ttttttttac aggggaaacg aaaatatatt
      361 tttcaaaagt atcgtttacc gctagctcag tcctaggtac aatgagcaca gtggcagctg
      421 tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa
      481 ttttgtttaa gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta
      541 ctgaaacatc ttaatcatgc gccggtgttt taaatatggc gactattaag caggtcgcgt
      601 tagaggcagg cctgtcgaaa agtactgtct cccgcttcat cgcccagaat ggatatgtct
      661 cggatgaagc tcgtaaaaag attgaagaag caatcaaaaa gttaaatttt accccaaatc
      721 tttctgccca gtcattgaaa acaaagaaga accagttagt ggggttactg cttccagaca
      781 ttagcaaccc tttttttcca atgttagcca agggtgctga agagttcctt aaggaaaagg
      841 gctatcgcgt catgttaggc aacatcggcg agaatcacca atcagagcat gactatctta
      901 aggtattgtt gcagagcaac gcggccggta ttatttcaac tcacgatttc aaggaggagt
      961 ttcctgacct ggacattccc tcggtaatcg tggaccgtgt tggccataag agtaattacg
     1021 gggtgttctc agacaacgaa agcggcggtc gtcttgccgc aaaggtcatt gttgaggcgg
     1081 gcgcgaaaaa cgtggccgta gtgtcaggac cactgaatgc tactaacatt aacaaccgtt
     1141 tcaaggcgag tatcgcgtac cttaaggaga agaaagtgga ctttaaggct ttgtattcgc
     1201 agtcctacga tttcgaggag atccagaagg aagctcgtga ggtgttagcg aataaagaga
     1261 attttgattc catcattgcg ccttcggaca ttcacgcgat ggcctacatt cacgagattc
     1321 atcgtcttga taaaaaaatt ccggaggaaa tccagattat tggttatgac gacatccaga
     1381 tgagtcagtt tatttatccc gctttaagca ccattcacca gagtgcgtat cagatgggac
     1441 tggaggctgc ccagttaatc tacaagattg ccactgacca gccaatcgag aagagcaaaa
     1501 ttgaattgcc tgtacattat gtagcacgcg aaacgattcg ccagaaggga aaagagtaat
     1561 aatcactttc agccaaaaaa cttaagaccg ccggtcttgt ccactacctt gcagtaatgc
     1621 ggtggacagg atcggcggtt ttcttttctc ttctcaaagg ctaggtggag gctcagtgat
     1681 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1741 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1801 tggagttcag ttcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     1861 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     1921 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     1981 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     2041 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     2101 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     2161 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     2221 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     2281 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     2341 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     2401 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     2461 tcatcttatt aagtctgacg ctctattcaa caaagccgcc gtccatgggt agggggcttc
     2521 aaatcgtccg ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc
     2581 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2641 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2701 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2761 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2821 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2881 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2941 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3001 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3061 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3121 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3181 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3241 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3301 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt
     3361 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3421 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     3481 aacgtggctt tcccccgccg ctctagaact agtggatcca aataaaacga aaggctcagt
     3541 cgaaagactg ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga cgtccaggtt
     3601 gggatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac
     3661 cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat
     3721 ccgcacgcat ctggaataag gaagtgccat tccgcctgac ctggaccaaa acgaaaaaag
     3781 gggagcggtt tcccgctccc ctcttttctg gaatttggta ccgaggaatg aagcaggatt
     3841 attatttgta tagttcatcc atgccatgtg taatcccagc agctgttaca aactcaagaa
     3901 ggaccatgtg gtctctcttt tcgttgggat ctttcgaaag ggcagattgt gtggacaggt
     3961 aatggttgtc tggtaaaagg acagggccat cgccaattgg agtattttgt tgataatggt
     4021 ctgctagttg aacgcttcca tcttcaatgt tgtgtctaat tttgaagtta actttgattc
     4081 cattcttttg tttgtctgcc atgatgtata cattgtgtga gttatagttg tattccaatt
     4141 tgtgtccaag aatgtttcca tcttctttaa aatcaatacc ttttaactcg attctattaa
     4201 caagggtatc accttcaaat ttgacttcag cacgtgtctt gtagttcccg tcatctttga
     4261 aaaatatagt tctttcctgt acataacctt cgggcatggc actcttgaaa aagtcatgct
     4321 gtttcatatg atctgggtat ctcgcaaagc attgaagacc atacgcgaaa gtagtgacaa
     4381 gtgttggcca tggaacaggt agttttccag tagtgcaaat aaatttaagg gtaagttttc
     4441 cgtatgttgc atcaccttca ccctctccac tgacagaaaa tttgtgccca ttaacatcac
     4501 c
//
