LOCUS       sequence-200257-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mCerulean.
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
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             171..884
                     /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
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(201..229)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     824..844
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      925..1011
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1549..2499)
                     /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      2547..2769
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3261..3355)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3386..4180)
                     /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(3497..3516)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4107..4126
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4359..4377
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
       61 actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa cggtttccct
      121 ctacaaataa ttttgtttaa cttttctaga tttaagaagg agatatacat atgagtaaag
      181 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
      241 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
      301 ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactc
      361 tcacttgggg tgttcaatgc tttgcaagat acccagatca tatgaaacag catgactttt
      421 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      481 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
      541 agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca
      601 acgctatctc agacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta
      661 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
      721 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
      781 aatctaagct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
      841 taacagctgc tgggattaca catggcatgg atgaactata caaataaatg tccagacctg
      901 caggcatgca agctctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
      961 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc
     1021 tagacctagg gtacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt
     1081 tatcagaatc gcagatccgg cttcagccgg tttgccggct gaaagcgcta tttcttccag
     1141 aattgccatg attttttccc cacgggaggc gtcactggct cccgtgttgt cggcagcttt
     1201 gattcgataa gcagcatcgc ctgtttcagg ctgtctatgt gtgactgttg agctgtaaca
     1261 agttgtctca ggtgttcaat ttcatgttct agttgctttg ttttactggt ttcacctgtt
     1321 ctattaggtg ttacatgctg ttcatctgtt acattgtcga tctgttcatg gtgaacagct
     1381 ttgaatgcac caaaaactcg taaaagctct gatgtatcta tcttttttac accgttttca
     1441 tctgtgcata tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg
     1501 tttgttagtc ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc
     1561 gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga
     1621 accattgaga tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag
     1681 ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga
     1741 atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag
     1801 ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg
     1861 gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat
     1921 tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa
     1981 catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt
     2041 tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac
     2101 atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc
     2161 actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat
     2221 ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt
     2281 tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg
     2341 gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag
     2401 tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat
     2461 cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg
     2521 attttaatca ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct
     2581 ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
     2641 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
     2701 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
     2761 gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt
     2821 tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc
     2881 gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt
     2941 ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact
     3001 acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca
     3061 cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg
     3121 ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg
     3181 tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc
     3241 ttactgtccc tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     3301 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     3361 gagctctcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     3421 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     3481 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     3541 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     3601 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     3661 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     3721 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     3781 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     3841 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     3901 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     3961 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     4021 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4081 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4141 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     4201 tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     4261 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccgacg
     4321 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4381 ttcgtcttca c
//
