LOCUS       sequence-152154-        2982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive expression of mcherry-pdt3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2982)
  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 2982)
  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: pZE27MC3
FEATURES             Location/Qualifiers
     source          1..2982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..798
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(236..254)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     504..523
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      925..1011
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1176..1764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1256..1275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1852..1946)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1977..2771)
                     /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(2088..2107)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2698..2717
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2950..2968
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
       61 agggaattca ttaaagagga caaaggtacc atggtgagca agggcgagga ggataacatg
      121 gccatcatca aggagttcat gcgcttcaag gttcacatgg agggctccgt gaacggccac
      181 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      241 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
      301 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
      361 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
      421 accgtgaccc aggactcctc cctgcaagac ggcgagttca tctacaaggt gaagctgcgc
      481 ggcaccaact tcccctccga cggccccgta atgcagaaga agactatggg ctgggaggcc
      541 tcctccgagc ggatgtaccc cgaggacggc gcgctgaagg gcgagatcaa gcagaggctg
      601 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
      661 cccgtgcaac tgcccggcgc gtacaacgtc aacatcaagt tggacatcac ctcccacaac
      721 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
      781 atggacgagc tgtacaaggc ggcgaacaaa aacgaagaaa acaccaacga agtgccgacc
      841 tttatgctga acgcgggcca ggcgaacaga agacgagttt aataagcttg atgggggatc
      901 ccatggtacg cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      961 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc
     1021 tagacctagg gcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     1621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctagtgcttg gattctcacc
     1861 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1921 cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgctcag
     1981 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     2041 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     2101 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     2161 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     2221 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     2281 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     2341 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     2401 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     2461 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     2521 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     2581 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     2641 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     2701 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     2761 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc
     2821 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     2881 ccagagggcg ccccagctgg caattccgac gtctaagaaa ccattattat catgacatta
     2941 acctataaaa ataggcgtat cacgaggccc tttcgtcttc ac
//
