LOCUS       sequence-445455-        4474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in response to L-Sorbose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4474)
  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 4474)
  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_LSorbose
FEATURES             Location/Qualifiers
     source          1..4474
                     /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(1687..1703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1687..1703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1700..1722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1883..2428)
                     /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(1963..1982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2539..3354)
                     /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(2539..3354)
                     /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     3262..3281
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3346..3365
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3475..3491
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3475..3491
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3492..3538
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3652..3674
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3666..3683
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3667..3683
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(3818..4474,1..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(3858..3878)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     join(4473..4474,1..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 atggggctga tgcgtctcct tcctaccttc
      181 acaaatgtta aatgtatgtt gttacctgtt tcacttttga ttataattgg aatgtaaccg
      241 gttacaagaa gaccgatcaa taatttcaca tttgtgattt ttgaggtgac aatatttctt
      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 taaatatgtc taatctgccg aagcgctacg
      601 accgcgcaac attagtaaaa atctcagact tgtattatat gcacggactt attcaacagg
      661 aaattagcga tatcgctcat attcatcgta ccgaaatttc ccgcattttg aaggcggctc
      721 gtgacgaggg tgttgttagt attgcgatta accccgaaac cactgccgta agccagttag
      781 tagacttttt caagcaaaag tacggtttac gtgatgcggt catcgttcca gctagcgaaa
      841 acggcggcaa tgaactgaat gcgttatctg tgtatgcttc tatgttctta tctaaaatca
      901 tcaaatctga cgatgtgatc ggattgtcat ggggcagcac tctgagttct gtcattagtc
      961 aattcccgac tgataaaaaa ttacacgaca tcaaggttgt gcccttagtt ggtggaccta
     1021 tgggccgcat cccgagcaac tatcacgtta attacttaac acatcgtctt gcgaatcgtt
     1081 tgaatgggac agccttcgta ctggactctc cggcctttgt acgctcaaaa gccttgcgca
     1141 aagaattgct ggctaaccca aatacacagg aaattcttag tttgtggacg cgtgttaaca
     1201 ttgccatttt cggtatgggg tcatcattga ttacggattc acccgactgg caagcttttt
     1261 atgaaaacac gaacttcaag tcatatttct cggctgagat ggtaggggac atcctttcac
     1321 atccgtttga taaatccggc aaagtaaccc gtgatatcga tagcattctg gtggcattgc
     1381 cattaagcgc attaaaaaag gtgccatact ctgttggcat cgcttttggc gaggataaag
     1441 ttacggcaat tctggctgcc ttgcgcggag gattgctgaa cactctgatt acgactgagg
     1501 ccaccgcaaa gtcgattaaa gatctgagct aataatcact ttcagccaaa aaacttaaga
     1561 ccgccggtct tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt
     1621 ctcttctcaa aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt
     1681 gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat
     1741 tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg agcggaaatg
     1801 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     1861 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     1921 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1981 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     2041 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     2101 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     2161 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     2221 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     2281 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     2341 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     2401 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaagtctg acgctctatt
     2461 caacaaagcc gccgtccatg ggtagggggc ttcaaatcgt ccgctctgcc agtgttacaa
     2521 ccaattaaca aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2581 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2641 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2701 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2761 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     2821 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2881 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     2941 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3001 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3061 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3121 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3181 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3241 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3301 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     3361 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3421 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcccccg ccgctctaga
     3481 actagtggat ccaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     3541 gttgtttgtc gcattatacg agacgtccag gttgggatac ctgaaacaaa acccatcgta
     3601 cggccaagga agtctccaat aactgtgatc caccacaagc gccagggttt tcccagtcac
     3661 gacgttgtaa aacgacggcc agtcatgcat aatccgcacg catctggaat aaggaagtgc
     3721 cattccgcct gacctggacc aaaacgaaaa aaggggagcg gtttcccgct cccctctttt
     3781 ctggaatttg gtaccgagga atgaagcagg attattattt gtatagttca tccatgccat
     3841 gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg
     3901 gatctttcga aagggcagat tgtgtggaca ggtaatggtt gtctggtaaa aggacagggc
     3961 catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgctt ccatcttcaa
     4021 tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct gccatgatgt
     4081 atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt
     4141 taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca aatttgactt
     4201 cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac
     4261 cttcgggcat ggcactcttg aaaaagtcat gctgtttcat atgatctggg tatctcgcaa
     4321 agcattgaag accatacgcg aaagtagtga caagtgttgg ccatggaaca ggtagttttc
     4381 cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc
     4441 cactgacaga aaatttgtgc ccattaacat cacc
//
