LOCUS       sequence-232729-        3058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses GFP fused with silaffin-R5 silica affinity 
            peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3058)
  AUTHORS   Olmez TT, Yuca E, Eyupoglu E, Catalak HB, Sahin O, Seker UOS
  TITLE     Autonomous Synthesis of Fluorescent Silica Biodots Using Engineered 
            Fusion Proteins.
  JOURNAL   ACS Omega. 2018 Jan 31;3(1):585-594. doi: 10.1021/acsomega.7b01769. 
            Epub 2018 Jan 18.
  PUBMED    30023783
REFERENCE   2  (bases 1 to 3058)
  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/acsomega.7b01769"; journalName: "ACS Omega"; date: 
            "2018-01-31- 31"; volume: "3"; issue: "1"; pages: "585-594"
COMMENT     Sequence Label: pZA-GFP-R5
FEATURES             Location/Qualifiers
     source          1..3058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..19,20..25,26..42,43..48,49..74)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             109..120
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..840
                     /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             127..840
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(157..185)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     780..800
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      933..1019
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1054..1071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1188..1733)
                     /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(1268..1287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1847..1941)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1962..2621)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2536..2555
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2622..2724)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    2763..2781
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2788..2806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
       61 aggacgcact gaccgaattc taccattcac ctcttggatt tgggtattaa agaggagaaa
      121 ggtaccatgc gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      181 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
      241 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
      301 acacttgtca ctactttcgg ttatggtgtt caatgctttg cgagataccc agatcatatg
      361 aaacagcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
      421 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
      481 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
      541 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
      601 aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta
      661 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
      721 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
      781 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
      841 ggtggcggtg gcagcagcag caaaaaaagc ggcagctata gcggcagcaa aggcagcaaa
      901 cgccgcattc tgtaaacgcg tgctagaggc atcaaataaa acgaaaggct cagtcgaaag
      961 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     1021 cgccgcccta gacctaggga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     1081 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     1141 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     1201 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     1261 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     1321 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     1381 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     1441 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     1501 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     1561 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     1621 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     1681 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     1741 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag
     1801 cacctgaagt cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa
     1861 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     1921 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     1981 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     2041 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     2101 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     2161 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     2221 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     2281 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     2341 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     2401 cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     2461 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     2521 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     2581 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     2641 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     2701 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtcaaagggc
     2761 cctccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacagtc
     2821 gacacccaaa tccaggaggt gattggtagt ggtggttaat gaaaattaac ttactactac
     2881 catatatctc tagatgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     2941 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3001 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc ttctcgag
//
