LOCUS       sequence-146491-        2988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive expression of gfp-pdt3E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2988)
  AUTHORS   Cameron DE, Collins JJ
  TITLE     Tunable protein degradation in bacteria.
  JOURNAL   Nat Biotechnol. 2014 Dec;32(12):1276-81. doi: 10.1038/nbt.3053. Epub
            2014 Nov 17.
  PUBMED    25402616
REFERENCE   2  (bases 1 to 2988)
  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: "10.1038/nbt.3053"; 
            journalName: "Nat Biotechnol"; date: "2014-12"; volume: "32"; issue:
            "12"; pages: "1276-81"
COMMENT     Sequence Label: pZE27GFP3E
FEATURES             Location/Qualifiers
     source          1..2988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..804
                     /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             91..804
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(121..149)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     744..764
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      931..1017
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1182..1770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1262..1281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1858..1952)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1983..2777)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2094..2113)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2704..2723
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2956..2974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
       61 agggaattca ttaaagagga caaaggtacc atgcgtaaag gagaagaact tttcactgga
      121 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
      181 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
      241 ggaaaactac ctgttccatg gccaacactt gtcactactt tcggttatgg tgttcaatgc
      301 tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa
      361 ggttacgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
      421 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
      481 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
      541 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
      601 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
      661 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
      721 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
      781 catggcatgg atgaactata caaagcggcg aacaaaaacg aagaaaacac caacgaagtg
      841 cccgacggaa tgctgaacgc gggccaggcg aacagaagac gagtttaata agcttgatgg
      901 gggatcccat ggtacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac
      961 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     1021 ccgccctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1621 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgactag tgcttggatt
     1861 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     1921 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
     1981 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     2041 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     2101 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     2161 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     2221 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     2281 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     2341 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     2401 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     2461 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     2521 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     2581 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     2641 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     2701 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     2761 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     2821 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     2881 accttaccag agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg
     2941 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
