LOCUS       sequence-200318-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mEmerald.
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-mEmerald
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             203..916
                     /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
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             203..916
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(233..261)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     856..876
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      957..1043
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1581..2531)
                     /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      2579..2801
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3293..3387)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3418..4212)
                     /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(3529..3548)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4139..4158
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4391,1..18)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ataggcgtat cacgaggccc tttcgtcttc accacagcta acaccacgtc gtccctatct
       61 gctgccctag gtctatgagt ggttgctgga taactttacg ggcatgcata aggctcgtat
      121 gatatattca gggagaccac aacggtttcc ctctacaaat aattttgttt aacttttcta
      181 gatttaagaa ggagatatac atatgagtaa aggagaagaa cttttcactg gagttgtccc
      241 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      301 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      361 acctgttcca tggccaacac ttgtcactac tctgacttat ggtgttcaat gctttgcaag
      421 atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg aaggttatgt
      481 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa
      541 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
      601 tggaaacatt cttggacaca aattggaata caactataac tcacacaagg tatacatcac
      661 ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa actagacaca acattgaaga
      721 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
      781 ccttttacca gacaaccatt acctgtccac acaatctaag ctttcgaaag atcccaacga
      841 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
      901 ggatgaacta tacaaataaa tgtccagacc tgcaggcatg caagctctag aggcatcaaa
      961 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1021 acgctctcct gagtaggaca aatccgccgc cctagaccta gggtacgggt tttgctgccc
     1081 gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc
     1141 ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag
     1201 gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca
     1261 ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt
     1321 ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg
     1381 ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct
     1441 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     1501 atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     1561 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     1621 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg
     1681 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     1741 agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg
     1801 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     1861 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     1921 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     1981 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     2041 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     2101 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     2161 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt
     2221 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     2281 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     2341 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     2401 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     2461 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     2521 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     2581 gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc
     2641 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     2701 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     2761 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     2821 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     2881 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     2941 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     3001 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     3061 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     3121 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     3181 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     3241 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac
     3301 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     3361 tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgctca
     3421 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3481 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     3541 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     3601 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     3661 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     3721 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     3781 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     3841 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     3901 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     3961 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4021 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     4081 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     4141 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     4201 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
     4261 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     4321 accagagggc gccccagctg gcaattccga cgtctaagaa accattatta tcatgacatt
     4381 aacctataaa a
//
