LOCUS       sequence-200540-        3932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding 2ndVal Katushka9-5.
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 Kat9-5
FEATURES             Location/Qualifiers
     source          1..3932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1023..1069
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1242..2192)
                     /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      2240..2462
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2968..3002)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3043..3837)
                     /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(3154..3173)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3764..3783
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcttcccaac cttaccagag ggcgccccag ctggcaattc cgacgtctaa gaaaccatta
       61 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaccaca
      121 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
      181 tacgggcatg cataaggctc gtatgatata ttcagggaga ccacaacggt ttccctctac
      241 aaataatttt gtttaacttt tctagattta agaaggagat atacatatgg tgtctgagct
      301 gattagcgag aacatgcaca tgaagctgta catggagggc accgtgaacg accaccactt
      361 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgaagatcaa
      421 ggtggtcgag ggcggccctt tacccttcgc cttcgacatc ctggctacca gcttcatgta
      481 cggcagcaag accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt
      541 ccctgagggc ttcacatggg agagaatcac cacatacgaa gacgggggcg tgctgaccgc
      601 tacccaggac accagcctgc agaacggctg cctgatctac aacgtcaaga tcaacggggt
      661 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctgggctggg aggccagcac
      721 cgagatgctg taccccgctg acagcggcct gaggggccac agccagatgg ccctgaagtt
      781 agtgggcggg ggctacctgc actgcagctt gaagaccaca tacagatcca agaaacccgc
      841 taagaacttg aagatgcccg gcttctactt cgtggacagg aaactggaaa gaatcaagga
      901 ggccgacaaa gagacctacg tcgagcagca cgagatggct gtggccaggt actgcgacct
      961 gcctagcaaa ctggggcaca gctaaatgtc cagacctgca ggcatgcaag ctctagaggc
     1021 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgaaatg
     1081 caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc
     1141 atatggacag ttttcccttt gatatctaac ggtgaacagt tgttctactt ttgtttgtta
     1201 gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta
     1261 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     1321 agatcatgct tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     1381 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta ggaatctgat
     1441 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     1501 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     1561 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     1621 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     1681 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     1741 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     1801 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     1861 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag
     1921 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     1981 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     2041 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     2101 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     2161 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     2221 tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt
     2281 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     2341 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     2401 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     2461 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     2521 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     2581 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     2641 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     2701 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
     2761 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
     2821 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
     2881 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt
     2941 caaccagacc cgccaggatt ctcatgcata aaaaacgccc ggcggcaacc gagcgttctg
     3001 aattaattaa tcatcgcgac tgcagagatc ccagagtcct tattagaaga actcgtcaag
     3061 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3121 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3181 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3241 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     3301 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3361 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3421 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3481 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     3541 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     3601 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     3661 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     3721 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     3781 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     3841 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     3901 cggcaagaaa gccatccagt ttactttgca gg
//
