LOCUS       sequence-200545-        3929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding Katushka-9-5.
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-9-5
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      904..950
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1123..2073)
                     /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      2121..2343
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2849..2883)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2924..3718)
                     /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(3035..3054)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3645..3664
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3897..3915
                     /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 atgtctgagc
      181 tgattagcga gaacatgcac atgaagctgt acatggaggg caccgtgaac gaccaccact
      241 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgaagatca
      301 aggtggtcga gggcggccct ttacccttcg ccttcgacat cctggctacc agcttcatgt
      361 acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
      421 tccctgaggg cttcacatgg gagagaatca ccacatacga agacgggggc gtgctgaccg
      481 ctacccagga caccagcctg cagaacggct gcctgatcta caacgtcaag atcaacgggg
      541 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actgggctgg gaggccagca
      601 ccgagatgct gtaccccgct gacagcggcc tgaggggcca cagccagatg gccctgaagt
      661 tagtgggcgg gggctacctg cactgcagct tgaagaccac atacagatcc aagaaacccg
      721 ctaagaactt gaagatgccc ggcttctact tcgtggacag gaaactggaa agaatcaagg
      781 aggccgacaa agagacctac gtcgagcagc acgagatggc tgtggccagg tactgcgacc
      841 tgcctagcaa actggggcac agctaaatgt ccagacctgc aggcatgcaa gctctagagg
      901 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgaaat
      961 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     1021 catatggaca gttttccctt tgatatctaa cggtgaacag ttgttctact tttgtttgtt
     1081 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     1141 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     1201 gagatcatgc ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     1261 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga
     1321 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     1381 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     1441 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     1501 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     1561 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     1621 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     1681 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     1741 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     1801 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     1861 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     1921 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     1981 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     2041 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     2101 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt
     2161 tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     2221 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     2281 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     2341 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     2401 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     2461 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     2521 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     2581 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     2641 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
     2701 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
     2761 agactggcta atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg
     2821 tcaaccagac ccgccaggat tctcatgcat aaaaaacgcc cggcggcaac cgagcgttct
     2881 gaattaatta atcatcgcga ctgcagagat cccagagtcc ttattagaag aactcgtcaa
     2941 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3001 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     3061 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     3121 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     3181 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     3241 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     3301 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     3361 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     3421 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     3481 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     3541 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     3601 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     3661 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     3721 gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg
     3781 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     3841 cagctggcaa ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3901 ggcgtatcac gaggcccttt cgtcttcac
//
