LOCUS       sequence-200538-        3932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding 2ndVal Katushka.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3932)
  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 3932)
  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: pEB2-2ndVal Katushka
FEATURES             Location/Qualifiers
     source          1..3932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..71
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      992..1038
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1211..2161)
                     /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      2209..2431
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2937..2971)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3012..3806)
                     /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(3123..3142)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3733..3752
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tggcaattcc gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
       61 tatcacgagg ccctttcgtc ttcaccacag ctaacaccac gtcgtcccta tctgctgccc
      121 taggtctatg agtggttgct ggataacttt acgggcatgc ataaggctcg tatgatatat
      181 tcagggagac cacaacggtt tccctctaca aataattttg tttaactttt ctagatttaa
      241 gaaggagata tacatatggt gtctgtgctg attaccgaga acatgcacat gaagctgtac
      301 atggagggca ccgtgaacga ccaccacttc aagtgcacat ccgagggcga aggcaagccc
      361 tacgagggca cccagaccat gaagatcaag gtggtcgagg gcggcccttt acccttcgcc
      421 ttcgacatcc tggctaccag cttcatgtac ggcagcaaga ccttcatcaa ccacacccag
      481 ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga gagaatcacc
      541 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctgca gaacggctgc
      601 ctgatctaca acgtcaagat caacggggtg aacttcccat ccaacggccc tgtgatgcag
      661 aagaaaacac tgggctggga ggccagcacc gagatgctgt accccgctga cagcggcctg
      721 aggggccaca gccagatggc cctgaagtta gtgggcgggg gctacctgca ctgcagcttg
      781 aagaccacat acagatccaa gaaacccgct aagaacttga agatgcccgg cttctacttc
      841 gtggacagga gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
      901 gagatggctg tggccaggta ctgcgacctg cctagcaaac tggggcacag ctaaatgtcc
      961 agacctgcag gcatgcaagc tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1021 ctgggccttt cgttttatct gttgaaatgc accaaaaact cgtaaaagct ctgatgtatc
     1081 tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg atatctaacg
     1141 gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc
     1201 cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt
     1261 tttgcgtgag ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa
     1321 attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc
     1381 ttagtccgtt acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca
     1441 tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt
     1501 ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt
     1561 aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat
     1621 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
     1681 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
     1741 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
     1801 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
     1861 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
     1921 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
     1981 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
     2041 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
     2101 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
     2161 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
     2221 gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg
     2281 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
     2341 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
     2401 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
     2461 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
     2521 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
     2581 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     2641 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     2701 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     2761 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     2821 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     2881 atcatcaaca ggcttacccg tcttactgtc aaccagaccc gccaggattc tcatgcataa
     2941 aaaacgcccg gcggcaaccg agcgttctga attaattaat catcgcgact gcagagatcc
     3001 cagagtcctt attagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3061 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3121 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     3181 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     3241 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     3301 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     3361 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3421 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3481 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3541 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     3601 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     3661 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     3721 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     3781 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     3841 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     3901 ggcttcccaa ccttaccaga gggcgcccca gc
//
