LOCUS       sequence-200530-        3914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding 2ndVal DsRed-Express2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3914)
  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 3914)
  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 DsRedEx2
FEATURES             Location/Qualifiers
     source          1..3914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1026..1072
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1245..2195)
                     /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      2243..2465
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2971..3005)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3046..3840)
                     /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(3157..3176)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3767..3786
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
       61 ccgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      121 ggccctttcg tcttcaccac agctaacacc acgtcgtccc tatctgctgc cctaggtcta
      181 tgagtggttg ctggataact ttacgggcat gcataaggct cgtatgatat attcagggag
      241 accacaacgg tttccctcta caaataattt tgtttaactt ttctagattt aagaaggaga
      301 tatacatatg gtggacagca ccgagaatgt gattaaaccg ttcatgcgct tcaaagttca
      361 catggagggt tcggttaacg gtcacgagtt cgagatcgaa ggcgaaggcg agggcaagcc
      421 gtatgaaggc actcaaaccg cgaaactgca agtgactaaa ggcggcccgc tgccttttgc
      481 gtgggacatc ctgagcccgc aatttcagta cggttctaaa gtgtatgtta aacacccagc
      541 ggatatcccg gactataaaa agctgtcttt tccggaaggt ttcaagtggg aacgcgtaat
      601 gaattttgaa gatggtggtg tcgtgaccgt cactcaggac tcctccctgc aggatggcac
      661 cttcatctat cacgttaaat ttattggtgt gaattttcca tctgatggcc cggtgatgca
      721 gaaaaagacg ttgggttggg agccgtctac cgaacgcctg tatccgcgcg atggtgtgct
      781 gaaaggcgaa attcataaag cgctgaaact gaaaggtggc ggccattatc tggtggaatt
      841 taaaagcatt tacatggcca agaaacctgt gaagctgcct ggctattact atgtggattc
      901 caaactggac atcacctctc ataatgaaga ttatacggtc gtagagcaat atgagcgcgc
      961 cgaggctcgt catcatctgt ttcagtaaat gtccagacct gcaggcatgc aagctctaga
     1021 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgaa
     1081 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     1141 tgcatatgga cagttttccc tttgatatct aacggtgaac agttgttcta cttttgtttg
     1201 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     1261 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     1321 ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     1381 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct
     1441 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     1501 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     1561 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     1621 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     1681 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     1741 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     1801 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     1861 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     1921 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     1981 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     2041 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     2101 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     2161 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     2221 taatcactat accaattgag atgggctagt caatgataat tactagtcct tttcctttga
     2281 gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     2341 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     2401 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     2461 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     2521 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     2581 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     2641 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     2701 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     2761 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     2821 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     2881 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     2941 tgtcaaccag acccgccagg attctcatgc ataaaaaacg cccggcggca accgagcgtt
     3001 ctgaattaat taatcatcgc gactgcagag atcccagagt ccttattaga agaactcgtc
     3061 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     3121 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     3181 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     3241 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     3301 gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     3361 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     3421 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     3481 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     3541 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     3601 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg
     3661 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     3721 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     3781 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     3841 gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct
     3901 tggcggcaag aaag
//
