LOCUS       sequence-200408-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mVenus JBC.
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-mVenus JBC
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             243..956
                     /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
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             243..956
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(273..301)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     896..916
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      997..1083
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1621..2571)
                     /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      2619..2841
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3333..3427)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3458..4252)
                     /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(3569..3588)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4179..4198
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtcttc accacagcta acaccacgtc gtccctatct gctgccctag gtctatgagt
      121 ggttgctgga taactttacg ggcatgcata aggctcgtat gatatattca gggagaccac
      181 aacggtttcc ctctacaaat aattttgttt aacttttcta gatttaagaa ggagatatac
      241 atatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      301 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      361 gaaaacttac ccttaaattg atttgcacta ctggaaaact acctgttcca tggccaacac
      421 ttgtcactac tttgggttat ggtctgcaat gctttgcgag atacccagat catatgaaac
      481 agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
      541 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
      601 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
      661 aattggaata caactataac tcacacaatg tatacatcac ggcagacaaa caaaagaatg
      721 gaatcaaagc gaacttcaaa attagacaca acattgaaga tggaggtgtt caactagcag
      781 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
      841 acctgtccta ccaatctaag ctttcgaaag atcccaacga aaagagagac cacatggtcc
      901 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataaa
      961 tgtccagacc tgcaggcatg caagctctag aggcatcaaa taaaacgaaa ggctcagtcg
     1021 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     1081 aatccgccgc cctagaccta gggtacgggt tttgctgccc gcaaacgggc tgttctggtg
     1141 ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc
     1201 tatttcttcc agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt
     1261 gtcggcagct ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt
     1321 tgagctgtaa caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg
     1381 gtttcacctg ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca
     1441 tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt
     1501 acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt
     1561 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
     1621 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     1681 ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc
     1741 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     1801 atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     1861 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
     1921 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     1981 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     2041 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     2101 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     2161 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     2221 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     2281 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     2341 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     2401 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     2461 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     2521 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     2581 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     2641 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     2701 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     2761 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     2821 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     2881 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     2941 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     3001 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     3061 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     3121 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     3181 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     3241 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     3301 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     3361 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     3421 aggagtccaa gcgagctctc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     3481 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     3541 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     3601 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     3661 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     3721 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     3781 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     3841 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     3901 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     3961 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4021 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     4081 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     4141 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     4201 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     4261 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     4321 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     4381 gcaattccga c
//
