LOCUS       sequence-200374-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mEYFP.
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-mEYFP
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..26
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             211..924
                     /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
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(241..269)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     864..884
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      965..1051
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1589..2539)
                     /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      2587..2809
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3301..3395)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3426..4220)
                     /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(3537..3556)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4147..4166
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac cacagctaac accacgtcgt
       61 ccctatctgc tgccctaggt ctatgagtgg ttgctggata actttacggg catgcataag
      121 gctcgtatga tatattcagg gagaccacaa cggtttccct ctacaaataa ttttgtttaa
      181 cttttctaga tttaagaagg agatatacat atgagtaaag gagaagaact tttcactgga
      241 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
      301 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
      361 ggaaaactac ctgttccatg gccaacactt gtcactactt tcggctatgg tcttcaatgc
      421 tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa
      481 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
      541 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
      601 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
      661 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
      721 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
      781 ggccctgtcc ttttaccaga caaccattac ctgtcctatc aatctaagct ttcgaaagat
      841 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
      901 catggcatgg atgaactata caaataaatg tccagacctg caggcatgca agctctagag
      961 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1021 gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg gtacgggttt
     1081 tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc gcagatccgg
     1141 cttcagccgg tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc
     1201 cacgggaggc gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc
     1261 ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat
     1321 ttcatgttct agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg
     1381 ttcatctgtt acattgtcga tctgttcatg gtgaacagct ttgaatgcac caaaaactcg
     1441 taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt
     1501 tccctttgat atgtaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc
     1561 actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct
     1621 ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatacttac
     1681 tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag
     1741 catcgtgtag tgtttttctt agtccgttat gtaggtagga atctgatgta atggttgttg
     1801 gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt
     1861 gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca
     1921 ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt
     1981 taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa
     2041 ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact
     2101 cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta
     2161 tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact aattctaatt
     2221 tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag
     2281 gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat
     2341 aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg
     2401 tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa
     2461 ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga
     2521 aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat
     2581 tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta
     2641 aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg
     2701 ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga
     2761 aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt
     2821 cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac
     2881 cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc
     2941 cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct
     3001 gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt
     3061 catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt
     3121 atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct
     3181 gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc
     3241 acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg
     3301 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     3361 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctctcga accccagagt
     3421 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     3481 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     3541 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     3601 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     3661 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     3721 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     3781 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     3841 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     3901 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     3961 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     4021 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     4081 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4141 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4201 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat
     4261 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     4321 ccaaccttac cagagggcgc cccagctggc aattccgacg tctaagaaac cattattatc
     4381 atgacattaa c
//
