LOCUS       sequence-200455-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding comClover3.
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-comClover3
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..55
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      994..1080
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1618..2568)
                     /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      2616..2838
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3330..3424)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3455..4249)
                     /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(3566..3585)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4176..4195
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
       61 cgtcttcacc acagctaaca ccacgtcgtc cctatctgct gccctaggtc tatgagtggt
      121 tgctggataa ctttacgggc atgcataagg ctcgtatgat atattcaggg agaccacaac
      181 ggtttccctc tacaaataat tttgtttaac ttttctagat ttaagaagga gatatacata
      241 tgagcaaagg tgaagaactg ttcaccggcg ttgtgccaat tctggttgag ctggatggtg
      301 acgtgaatgg ccacaaattt tccgtgagag gtgaaggcga gggtgatgct actaatggca
      361 aactgactct gaaattcatc tgtaccaccg gcaaactgcc tgttccgtgg ccaactctgg
      421 tcactacttt cggttacggc gtggcttgtt tttcgcgtta cccggatcac atgaaacagc
      481 atgacttctt caaatctgcc atgccggaag gctatgtcca agaacgtacg atctctttca
      541 aggacgacgg cacctataaa acccgtgccg aagttaaatt cgagggtgac accctggtta
      601 accgcatcga actgaaaggc attgacttca aagaggacgg caacattctg ggtcacaagc
      661 tggaatacaa cttcaactcc cactacgttt acattactgc tgacaagcag aaaaactgca
      721 tcaaagcaaa cttcaagatc cgtcacaacg ttgaagatgg tagcgtacag ctggcagatc
      781 actaccagca gaacactcca atcggtgatg gcccagtact gctgccagat aaccattacc
      841 tgtcccacca gagcaaactg tctaaagacc cgaacgaaaa acgtgaccac atggtactgc
      901 tggaatttgt taccgcggca ggcattaccc acggtatgga cgaactgtat aaataaatgt
      961 ccagacctgc aggcatgcaa gctctagagg catcaaataa aacgaaaggc tcagtcgaaa
     1021 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1081 ccgccgccct agacctaggg tacgggtttt gctgcccgca aacgggctgt tctggtgttg
     1141 ctagtttgtt atcagaatcg cagatccggc ttcagccggt ttgccggctg aaagcgctat
     1201 ttcttccaga attgccatga ttttttcccc acgggaggcg tcactggctc ccgtgttgtc
     1261 ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg tgactgttga
     1321 gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt tttactggtt
     1381 tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat ctgttcatgg
     1441 tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca
     1501 ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg aacagttgtt
     1561 ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca
     1621 gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca
     1681 tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt ttgcctcaaa
     1741 actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttatg
     1801 taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt
     1861 gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt
     1921 atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc
     1981 tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc
     2041 aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt
     2101 cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa
     2161 agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa
     2221 tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca
     2281 ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag
     2341 ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg
     2401 agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt
     2461 ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata
     2521 gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg
     2581 gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt
     2641 ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt
     2701 aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat
     2761 tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc
     2821 cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca
     2881 aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct
     2941 cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc
     3001 gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta
     3061 aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa
     3121 agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac
     3181 tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc
     3241 cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc
     3301 ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     3361 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     3421 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     3481 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     3541 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     3601 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     3661 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     3721 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     3781 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     3841 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     3901 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     3961 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     4021 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     4081 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     4141 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     4201 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     4261 ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     4321 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     4381 attccgacgt c
//
