LOCUS       sequence-200316-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mGFPmut3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4391)
  AUTHORS   Balleza E, Kim JM, Cluzel P
  TITLE     Systematic characterization of maturation time of fluorescent 
            proteins in living cells.
  JOURNAL   Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 
            2017 Nov 20.
  PUBMED    29320486
REFERENCE   2  (bases 1 to 4391)
  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/nmeth.4509"; journalName: "Nat Methods"; date: "2018-01"; 
            volume: "15"; issue: "1"; pages: "47-51"
COMMENT     Sequence Label: pEB1-mGFPmut3
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             193..906
                     /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
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(223..251)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     846..866
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      947..1033
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1571..2521)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2569..2791
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3283..3377)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3408..4202)
                     /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(3519..3538)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4129..4148
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4381..4391,1..8)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cacgaggccc tttcgtcttc accacagcta acaccacgtc gtccctatct gctgccctag
       61 gtctatgagt ggttgctgga taactttacg ggcatgcata aggctcgtat gatatattca
      121 gggagaccac aacggtttcc ctctacaaat aattttgttt aacttttcta gatttaagaa
      181 ggagatatac atatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
      241 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
      301 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
      361 tggccaacac ttgtcactac tttcggttat ggtgttcaat gctttgcgag atacccagat
      421 catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga
      481 actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt
      541 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
      601 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
      661 caaaagaatg gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt
      721 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
      781 gacaaccatt acctgtccac acaatctaag ctttcgaaag atcccaacga aaagagagac
      841 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
      901 tacaaataaa tgtccagacc tgcaggcatg caagctctag aggcatcaaa taaaacgaaa
      961 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     1021 gagtaggaca aatccgccgc cctagaccta gggtacgggt tttgctgccc gcaaacgggc
     1081 tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg
     1141 ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag gcgtcactgg
     1201 ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca ggctgtctat
     1261 gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt ctagttgctt
     1321 tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg ttacattgtc
     1381 gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc
     1441 tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg
     1501 gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc
     1561 cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt
     1621 tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa
     1681 attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc
     1741 ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca
     1801 tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt
     1861 ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt
     1921 aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat
     1981 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
     2041 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
     2101 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
     2161 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
     2221 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
     2281 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
     2341 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
     2401 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
     2461 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
     2521 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
     2581 gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg
     2641 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
     2701 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
     2761 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
     2821 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
     2881 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
     2941 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     3001 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     3061 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     3121 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     3181 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     3241 atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa
     3301 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     3361 atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgctca gaagaactcg
     3421 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     3481 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     3541 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     3601 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     3661 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     3721 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     3781 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     3841 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     3901 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     3961 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     4021 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     4081 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     4141 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     4201 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     4261 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     4321 gccccagctg gcaattccga cgtctaagaa accattatta tcatgacatt aacctataaa
     4381 aataggcgta t
//
