LOCUS       sequence-200314-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mEGFP.
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-mEGFP
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             197..910
                     /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
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             197..910
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(227..255)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     850..870
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      951..1037
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1575..2525)
                     /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      2573..2795
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3287..3381)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3412..4206)
                     /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(3523..3542)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4133..4152
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4385..4391,1..12)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtatcacgag gccctttcgt cttcaccaca gctaacacca cgtcgtccct atctgctgcc
       61 ctaggtctat gagtggttgc tggataactt tacgggcatg cataaggctc gtatgatata
      121 ttcagggaga ccacaacggt ttccctctac aaataatttt gtttaacttt tctagattta
      181 agaaggagat atacatatga gtaaaggaga agaacttttc actggagttg tcccaattct
      241 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
      301 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
      361 tccatggcca acacttgtca ctactctgac ttatggtgtt caatgctttt caagataccc
      421 agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
      481 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga
      541 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa
      601 cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga
      661 caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag
      721 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
      781 accagacaac cattacctgt ccacacaatc taagctttcg aaagatccca acgaaaagag
      841 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
      901 actatacaaa taaatgtcca gacctgcagg catgcaagct ctagaggcat caaataaaac
      961 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1021 tcctgagtag gacaaatccg ccgccctaga cctagggtac gggttttgct gcccgcaaac
     1081 gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc agccggtttg
     1141 ccggctgaaa gcgctatttc ttccagaatt gccatgattt tttccccacg ggaggcgtca
     1201 ctggctcccg tgttgtcggc agctttgatt cgataagcag catcgcctgt ttcaggctgt
     1261 ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca tgttctagtt
     1321 gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca tctgttacat
     1381 tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg
     1441 tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt
     1501 aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa
     1561 gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt
     1621 tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc
     1681 aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt
     1741 tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca
     1801 ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca
     1861 acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg
     1921 ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac
     1981 tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct aatctctata
     2041 tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata aatcctcata
     2101 gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa tttttttaac
     2161 tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc gcttgagaac
     2221 ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt cctgatttcc
     2281 acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc attttcccta
     2341 ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg ttctttcctt
     2401 gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag cttggtttca
     2461 tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac aactaattca
     2521 gacatacatc tcaattggtc taggtgattt taatcactat accaattgag atgggctagt
     2581 caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt ctgctagacc
     2641 tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag acctttgtgt
     2701 gttttttttg tttatattca agtggttata atttatagaa taaagaaaga ataaaaaaag
     2761 ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt tccgcagtat
     2821 tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta aaaccctaaa
     2881 ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt tgtctccgac
     2941 catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc tcacggctct
     3001 ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg caaggaaact
     3061 acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct
     3121 atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg
     3181 accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag
     3241 cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa
     3301 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     3361 ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa
     3421 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     3481 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     3541 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     3601 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     3661 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     3721 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     3781 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     3841 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     3901 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     3961 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     4021 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4081 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4141 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4201 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     4261 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     4321 gggcgcccca gctggcaatt ccgacgtcta agaaaccatt attatcatga cattaaccta
     4381 taaaaatagg c
//
