LOCUS       sequence-200529-        3929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding Katushka.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3929)
  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 3929)
  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: pEB2-Katushka
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..76
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      994..1040
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1213..2163)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2211..2433
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2939..2973)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3014..3808)
                     /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(3125..3144)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3735..3754
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccagctggca attccgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
       61 aggcgtatca cgaggccctt tcgtcttcac cacagctaac accacgtcgt ccctatctgc
      121 tgccctaggt ctatgagtgg ttgctggata actttacggg catgcataag gctcgtatga
      181 tatattcagg gagaccacaa cggtttccct ctacaaataa ttttgtttaa cttttctaga
      241 tttaagaagg agatatacat atgtctgtgc tgattaccga gaacatgcac atgaagctgt
      301 acatggaggg caccgtgaac gaccaccact tcaagtgcac atccgagggc gaaggcaagc
      361 cctacgaggg cacccagacc atgaagatca aggtggtcga gggcggccct ttacccttcg
      421 ccttcgacat cctggctacc agcttcatgt acggcagcaa gaccttcatc aaccacaccc
      481 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagaatca
      541 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctg cagaacggct
      601 gcctgatcta caacgtcaag atcaacgggg tgaacttccc atccaacggc cctgtgatgc
      661 agaagaaaac actgggctgg gaggccagca ccgagatgct gtaccccgct gacagcggcc
      721 tgaggggcca cagccagatg gccctgaagt tagtgggcgg gggctacctg cactgcagct
      781 tgaagaccac atacagatcc aagaaacccg ctaagaactt gaagatgccc ggcttctact
      841 tcgtggacag gagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
      901 acgagatggc tgtggccagg tactgcgacc tgcctagcaa actggggcac agctaaatgt
      961 ccagacctgc aggcatgcaa gctctagagg catcaaataa aacgaaaggc tcagtcgaaa
     1021 gactgggcct ttcgttttat ctgttgaaat gcaccaaaaa ctcgtaaaag ctctgatgta
     1081 tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatctaa
     1141 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     1201 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     1261 tttttgcgtg agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa
     1321 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     1381 tcttagtccg ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     1441 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     1501 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     1561 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     1621 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     1681 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     1741 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     1801 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     1861 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     1921 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     1981 gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt
     2041 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     2101 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     2161 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     2221 atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     2281 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     2341 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     2401 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     2461 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     2521 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
     2581 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
     2641 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
     2701 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
     2761 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
     2821 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
     2881 gtatcatcaa caggcttacc cgtcttactg tcaaccagac ccgccaggat tctcatgcat
     2941 aaaaaacgcc cggcggcaac cgagcgttct gaattaatta atcatcgcga ctgcagagat
     3001 cccagagtcc ttattagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     3061 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     3121 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     3181 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     3241 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     3301 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     3361 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     3421 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     3481 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     3541 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     3601 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     3661 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     3721 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     3781 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     3841 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     3901 agggcttccc aaccttacca gagggcgcc
//
