LOCUS       sequence-147374-        2907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive expression of gfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2907)
  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 2907)
  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: pZE27GFP
FEATURES             Location/Qualifiers
     source          1..2907
                     /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"
     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      850..936
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1101..1689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1181..1200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1777..1871)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1902..2696)
                     /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(2013..2032)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2623..2642
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2875..2893
                     /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 ggttatgtac 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 caaataataa gcttgatggg ggatcccatg gtacgcgtgc
      841 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      901 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggcgtt
      961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1501 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     1561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1741 tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa aaaacgcccg
     1801 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     1861 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     1921 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     1981 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     2041 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     2101 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     2161 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     2221 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     2281 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     2341 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     2401 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     2461 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     2521 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     2581 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     2641 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     2701 aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca gatccttggc
     2761 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     2821 gctggcaatt ccgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2881 cgtatcacga ggccctttcg tcttcac
//
