LOCUS       sequence-445467-        4460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in response to 4-Vinylphenol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4460)
  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 4460)
  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_4Vinylphenol
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        452..501
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(1673..1689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1673..1689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1686..1708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1869..2414)
                     /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(1949..1968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2525..3340)
                     /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(2525..3340)
                     /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     3248..3267
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3332..3351
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3461..3477
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3461..3477
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3478..3524
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3638..3660
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3652..3669
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3653..3669
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(3804..4460,1..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(3844..3864)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     join(4459..4460,1..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 atggggctgt caattcctcc aaactgattt
      181 gttatctaat gagcatttga cggctaaatt caccattact ataatgaagt aaccgctatc
      241 acaccctcat cataacgggg gtatggtggt tagcaaacag acaatttggt tcggcaggcg
      301 aatcaatcgt gcggatggag ttgaaaaatt tcttgcaaaa aaaaaccccg ccgaagcggg
      361 gttttttttt ttacagggga aacgaaaata tatttttcaa aagtatcgtt taccgctagc
      421 tcagtcctag gtacaatgag cacagtggca gctgtcaccg gatgtgcttt ccggtctgat
      481 gagtccgtga ggacgaaaca gcctctacaa ataattttgt ttaagggccc aagttcactt
      541 aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa catcttaatc atgcgccggt
      601 gttttaaata tggaccttga tcgcttacag accttcctta aagtcgtaca atacggatcg
      661 tttcaacaag tcgcagcgcg tgagtaccgc agtcaacgta cagtctctaa gcagatgaca
      721 cagctggaaa atgagttaaa agtaactttg ttcgaccgcg gtcagaaccg catccaactt
      781 acgccgcagg gccgtttatt ctgggcctcc gctcaagaca tcgtgaataa ctacactacg
      841 gctcttacgg aattacgtca atttaatgtg ccaaccgacc aaattcttcg cgttggctat
      901 ttttcggcct tcgagcaacg tttattattg cccgcgttat atgatttaaa gcaacaacac
      961 tccgagttac agttagtagt gcgccagggc tctaacgagc atcttgcaca gcaagttgcc
     1021 gatggctccc ttgatttagc attatctatt aactatggac gcccagctgt aacccctgaa
     1081 tcccaactga ccgctgtacc tatttatcat aaccagatgg tgatcggggt ctcacgcctt
     1141 aacccattat cgcgtctttc acagttgccg ccatcggcgt tagcaacgct gcccatttta
     1201 tattactccc ctgagtcttc aactttcttg ctggaatcct tccttgcctc agctcctttc
     1261 atccaggatt atgagcaaat tcgccgtgtt tcttccgccg agcaaatgca ccttttagtg
     1321 gccttgaatc aagctttagc cttctatccg gctggcttgg tgccaaccaa acacgatgag
     1381 caagttgcgt atcttccgat taccgacgcc gctcagcagg gctacgacat tgtcgccttg
     1441 ttgaaatcta accgtagtcg ccctctgatc gccaagttgg tccagcgctt gaaggctaat
     1501 gccaaatccg atgaataata atcactttca gccaaaaaac ttaagaccgc cggtcttgtc
     1561 cactaccttg cagtaatgcg gtggacagga tcggcggttt tcttttctct tctcaaaggc
     1621 taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca tggtcatagc
     1681 tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg cgctatccga
     1741 caatctccaa gacattaggt ggagttcagt tcggcgagcg gaaatggctt acgaacgggg
     1801 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     1861 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
     1921 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
     1981 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     2041 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     2101 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     2161 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     2221 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     2281 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     2341 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     2401 caaaacgatc tcaagaagat catcttatta agtctgacgc tctattcaac aaagccgccg
     2461 tccatgggta gggggcttca aatcgtccgc tctgccagtg ttacaaccaa ttaacaaatt
     2521 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     2581 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     2641 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     2701 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2761 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     2821 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2881 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2941 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3001 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3061 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3121 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3181 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3241 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3301 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3361 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3421 atcagagatt ttgagacaca acgtggcttt cccccgccgc tctagaacta gtggatccaa
     3481 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcgcat
     3541 tatacgagac gtccaggttg ggatacctga aacaaaaccc atcgtacggc caaggaagtc
     3601 tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg
     3661 acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc
     3721 tggaccaaaa cgaaaaaagg ggagcggttt cccgctcccc tcttttctgg aatttggtac
     3781 cgaggaatga agcaggatta ttatttgtat agttcatcca tgccatgtgt aatcccagca
     3841 gctgttacaa actcaagaag gaccatgtgg tctctctttt cgttgggatc tttcgaaagg
     3901 gcagattgtg tggacaggta atggttgtct ggtaaaagga cagggccatc gccaattgga
     3961 gtattttgtt gataatggtc tgctagttga acgcttccat cttcaatgtt gtgtctaatt
     4021 ttgaagttaa ctttgattcc attcttttgt ttgtctgcca tgatgtatac attgtgtgag
     4081 ttatagttgt attccaattt gtgtccaaga atgtttccat cttctttaaa atcaatacct
     4141 tttaactcga ttctattaac aagggtatca ccttcaaatt tgacttcagc acgtgtcttg
     4201 tagttcccgt catctttgaa aaatatagtt ctttcctgta cataaccttc gggcatggca
     4261 ctcttgaaaa agtcatgctg tttcatatga tctgggtatc tcgcaaagca ttgaagacca
     4321 tacgcgaaag tagtgacaag tgttggccat ggaacaggta gttttccagt agtgcaaata
     4381 aatttaaggg taagttttcc gtatgttgca tcaccttcac cctctccact gacagaaaat
     4441 ttgtgcccat taacatcacc
//
