LOCUS       sequence-200299-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mCerulean ME.
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-mCerulean ME
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             130..843
                     /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
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(160..188)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     783..803
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      884..970
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1508..2458)
                     /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      2506..2728
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3220..3314)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3345..4139)
                     /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(3456..3475)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4066..4085
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4318..4336
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tatgagtggt tgctggataa ctttacgggc atgcataagg ctcgtatgat atattcaggg
       61 agaccacaac ggtttccctc tacaaataat tttgtttaac ttttctagat ttaagaagga
      121 gatatacata tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
      181 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
      241 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
      301 ccaacacttg tcactactct cacttggggt gttcaatgct ttgcaagata cccagatcat
      361 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
      421 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
      481 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
      541 ggacacaaat tggaatacaa cgctatctca gacaatgtat acatcacggc agacaaacaa
      601 aagaatggaa tcaaagctca cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
      661 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
      721 aaccattacc tgtccacaca atctaagctt tcgaaagatc ccaacgaaaa gagagaccac
      781 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
      841 aaataaatgt ccagacctgc aggcatgcaa gctctagagg catcaaataa aacgaaaggc
      901 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      961 taggacaaat ccgccgccct agacctaggg tacgggtttt gctgcccgca aacgggctgt
     1021 tctggtgttg ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg
     1081 aaagcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc
     1141 ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg
     1201 tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt
     1261 tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat
     1321 ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat
     1381 cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg
     1441 aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat
     1501 aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt
     1561 gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt
     1621 ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta
     1681 gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt
     1741 ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga
     1801 aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag
     1861 tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt
     1921 agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct
     1981 tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt
     2041 tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa
     2101 gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat
     2161 agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc
     2221 tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt
     2281 tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt
     2341 tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg
     2401 ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac
     2461 atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat
     2521 aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg
     2581 gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt
     2641 ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa
     2701 gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa
     2761 ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa
     2821 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
     2881 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
     2941 aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata
     3001 atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt
     3061 gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt
     3121 cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc
     3181 atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc
     3241 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     3301 tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca
     3361 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3421 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3481 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     3541 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     3601 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     3661 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     3721 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     3781 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     3841 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     3901 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     3961 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4021 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4081 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4141 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     4201 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     4261 ccagctggca attccgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4321 aggcgtatca cgaggccctt tcgtcttcac cacagctaac accacgtcgt ccctatctgc
     4381 tgccctaggt c
//
