LOCUS       sequence-200443-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding coCeruleanME.
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-coCeruleanME
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      973..1059
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1597..2547)
                     /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      2595..2817
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3309..3403)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3434..4228)
                     /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(3545..3564)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4155..4174
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacca cagctaacac
       61 cacgtcgtcc ctatctgctg ccctaggtct atgagtggtt gctggataac tttacgggca
      121 tgcataaggc tcgtatgata tattcaggga gaccacaacg gtttccctct acaaataatt
      181 ttgtttaact tttctagatt taagaaggag atatacatat gagcaagggc gaggagctgt
      241 ttactggtgt ggttcctatc ctggtcgaac tggacggtga tgtcaacggt cataagttca
      301 gcgttagcgg cgagggtgaa ggcgacgcaa cctacggtaa gctgaccctg aagttcattt
      361 gtacgacggg taagctgccg gtgccttggc ctaccctggt taccaccctg acctggggtg
      421 ttcaatgctt cgctcgctat ccagaccaca tgaagcagca cgattttttt aagagcgcca
      481 tgccagaggg ttacgttcag gagcgcacca ttttctttaa agatgatggt aattacaaga
      541 ctcgcgcaga ggtgaagttt gaaggcgaca ctctggtgaa tcgtattgag ctgaagggca
      601 tcgattttaa ggaagatggt aatatcctgg gccataaact ggaatataat gcgattagcg
      661 ataatgtgta tatcaccgcc gataaacaaa agaatggtat taaggcgcat tttaaaattc
      721 gccataatat cgaggacggc tctgtgcaac tggctgacca ttatcaacaa aataccccga
      781 ttggcgacgg tcctgttctg ctgccggaca atcactatct gagcactcaa tctaagctga
      841 gcaaagatcc taatgagaag cgcgatcaca tggtgctgct ggaatttgta acggctgctg
      901 gcatcacgca tggcatggat gagctgtata aataaatgtc cagacctgca ggcatgcaag
      961 ctctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1021 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggt
     1081 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
     1141 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
     1201 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
     1261 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
     1321 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
     1381 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
     1441 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
     1501 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     1561 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     1621 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     1681 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     1741 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     1801 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     1861 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     1921 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     1981 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     2041 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     2101 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     2161 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     2221 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
     2281 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     2341 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     2401 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     2461 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     2521 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     2581 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     2641 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     2701 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     2761 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     2821 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
     2881 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
     2941 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
     3001 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
     3061 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
     3121 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
     3181 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
     3241 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta
     3301 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     3361 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctctcgaac
     3421 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3481 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3541 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3601 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3661 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     3721 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     3781 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     3841 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     3901 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     3961 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     4021 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     4081 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     4141 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     4201 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     4261 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     4321 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc taagaaacca
     4381 ttattatcat g
//
