LOCUS       sequence-200406-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mVenus NB.
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 NB
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             202..915
                     /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
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             202..915
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(232..260)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     855..875
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      956..1042
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1580..2530)
                     /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      2578..2800
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3292..3386)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3417..4211)
                     /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(3528..3547)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4138..4157
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4390..4391,1..17)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taggcgtatc acgaggccct ttcgtcttca ccacagctaa caccacgtcg tccctatctg
       61 ctgccctagg tctatgagtg gttgctggat aactttacgg gcatgcataa ggctcgtatg
      121 atatattcag ggagaccaca acggtttccc tctacaaata attttgttta acttttctag
      181 atttaagaag gagatataca tatgagtaaa ggagaagaac ttttcactgg agttgtccca
      241 attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt
      301 gaaggtgatg caacatacgg aaaacttacc cttaaattga tttgcactac tggaaaacta
      361 cctgttccat ggccaacact tgtcactact ttgggttatg gtgttcaatg ctttgcgaga
      421 tacccagatc atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta
      481 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag
      541 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
      601 ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt atacatcacg
      661 gcagacaaac aaaagaatgg aatcaaagcg aacttcaaaa ttagacacaa cattgaagat
      721 ggaggtgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
      781 cttttaccag acaaccatta cctgtcctac caatctaagc tttcgaaaga tcccaacgaa
      841 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
      901 gatgaactat acaaataaat gtccagacct gcaggcatgc aagctctaga ggcatcaaat
      961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1021 cgctctcctg agtaggacaa atccgccgcc ctagacctag ggtacgggtt ttgctgcccg
     1081 caaacgggct gttctggtgt tgctagtttg ttatcagaat cgcagatccg gcttcagccg
     1141 gtttgccggc tgaaagcgct atttcttcca gaattgccat gattttttcc ccacgggagg
     1201 cgtcactggc tcccgtgttg tcggcagctt tgattcgata agcagcatcg cctgtttcag
     1261 gctgtctatg tgtgactgtt gagctgtaac aagttgtctc aggtgttcaa tttcatgttc
     1321 tagttgcttt gttttactgg tttcacctgt tctattaggt gttacatgct gttcatctgt
     1381 tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc gtaaaagctc
     1441 tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt ttccctttga
     1501 tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt cactgataga
     1561 tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc tctagtgtgg
     1621 ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta ctttgcatgt
     1681 cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa gcatcgtgta
     1741 gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt ggtattttgt
     1801 caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt tgtctatcta
     1861 gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc accaatttca
     1921 tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg ttaagccttt
     1981 taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca aggctaatct
     2041 ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac tcataaatcc
     2101 tcatagagta tttgttttca aaagacttaa catgttccag attatatttt atgaattttt
     2161 ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat ttttcgcttg
     2221 agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa ggattcctga
     2281 tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca taagcatttt
     2341 ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc gtccgttctt
     2401 tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa attagcttgg
     2461 tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg aaaacaacta
     2521 attcagacat acatctcaat tggtctaggt gattttaatc actataccaa ttgagatggg
     2581 ctagtcaatg ataattacta gtccttttcc tttgagttgt gggtatctgt aaattctgct
     2641 agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt
     2701 tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa
     2761 aaaagataaa aagaatagat cccagccctg tgtataactc actactttag tcagttccgc
     2821 agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc
     2881 ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct
     2941 ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg
     3001 gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg
     3061 aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt
     3121 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
     3181 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
     3241 ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc
     3301 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     3361 cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgctcag
     3421 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     3481 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     3541 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     3601 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     3661 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     3721 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     3781 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     3841 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     3901 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     3961 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     4021 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     4081 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     4141 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     4201 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc
     4261 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     4321 ccagagggcg ccccagctgg caattccgac gtctaagaaa ccattattat catgacatta
     4381 acctataaaa a
//
