LOCUS       sequence-200456-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding coClover.
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-coClover
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..23
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      962..1048
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1586..2536)
                     /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      2584..2806
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3298..3392)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3423..4217)
                     /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(3534..3553)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4144..4163
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 taaaaatagg cgtatcacga ggccctttcg tcttcaccac agctaacacc acgtcgtccc
       61 tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct
      121 cgtatgatat attcagggag accacaacgg tttccctcta caaataattt tgtttaactt
      181 ttctagattt aagaaggaga tatacatatg agcaaaggtg aagaactgtt caccggcgtt
      241 gtgccaattc tggttgagct ggatggtgac gtgaatggcc acaaattttc cgtgagaggt
      301 gaaggcgagg gtgatgctac taatggcaaa ctgactctga aattcatctg taccaccggc
      361 aaactgcctg ttccgtggcc aactctggtc actactttcg gttacggcgt ggcttgtttt
      421 tcgcgttacc cggatcacat gaaacagcat gacttcttca aatctgccat gccggaaggc
      481 tatgtccaag aacgtacgat ctctttcaag gacgacggca cctataaaac ccgtgccgaa
      541 gttaaattcg agggtgacac cctggttaac cgcatcgaac tgaaaggcat tgacttcaaa
      601 gaggacggca acattctggg tcacaagctg gaatacaact tcaactccca caacgtttac
      661 attactgctg acaagcagaa aaacggcatc aaagcaaact tcaagatccg tcacaacgtt
      721 gaagatggta gcgtacagct ggcagatcac taccagcaga acactccaat cggtgatggc
      781 ccagtactgc tgccagataa ccattacctg tcccaccaga gcgcactgtc taaagacccg
      841 aacgaaaaac gtgaccacat ggtactgctg gaatttgtta ccgcggcagg cattacccac
      901 ggtatggacg aactgtataa ataaatgtcc agacctgcag gcatgcaagc tctagaggca
      961 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1021 ggtgaacgct ctcctgagta ggacaaatcc gccgccctag acctagggta cgggttttgc
     1081 tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt
     1141 cagccggttt gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac
     1201 gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg
     1261 tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc
     1321 atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc
     1381 atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa
     1441 aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc
     1501 ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact
     1561 gatagataca agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta
     1621 gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt
     1681 gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat
     1741 cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta
     1801 ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc
     1861 tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca
     1921 atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa
     1981 gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc
     2041 taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat
     2101 aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga
     2161 atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt
     2221 cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat
     2281 tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag
     2341 cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc
     2401 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
     2461 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
     2521 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
     2581 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
     2641 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
     2701 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
     2761 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
     2821 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
     2881 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
     2941 ttgtctccga ccatcaggca cctgagtcgc trtctttttc gtgacattca gttcgctgcg
     3001 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
     3061 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
     3121 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
     3181 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
     3241 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt
     3301 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     3361 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc
     3421 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     3481 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     3541 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     3601 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     3661 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     3721 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     3781 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     3841 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     3901 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     3961 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     4021 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     4081 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     4141 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     4201 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     4261 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     4321 accttaccag agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg
     4381 acattaacct a
//
