LOCUS       sequence-201141-        3911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding E2-Crimson.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3911)
  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 3911)
  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-E2-Crimson
FEATURES             Location/Qualifiers
     source          1..3911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      886..932
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1105..2055)
                     /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      2103..2325
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2831..2865)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2906..3700)
                     /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(3017..3036)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3627..3646
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3879..3897
                     /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 atggacagca
      181 ccgagaatgt gattaaaccg ttcatgcgct tcaaagttca catggagggt tcggttaacg
      241 gtcacgagtt cgagatcgaa ggcgtaggcg agggcaagcc gtatgaaggc actcaaaccg
      301 cgaaactgca agtgactaaa ggcggcccgc tgccttttgc gtgggacatc ctgagcccgc
      361 aatttttcta cggttctaaa gcatatatca aacacccagc ggatatcccg gactatttaa
      421 agcagtcttt tccggaaggt ttcaagtggg aacgcgtaat gaattttgaa gatggtggtg
      481 tcgtgaccgt cactcaggac tcctccctgc aggatggcac cttgatctat cacgttaaat
      541 ttattggtgt gaattttcca tctgatggcc cggtgatgca gaaaaagacg ttgggttggg
      601 agccgtctac cgaacgcaac tatccgcgcg atggtgtgct gaaaggcgaa aatcatatgg
      661 cgctgaaact gaaaggtggc ggccattatc tgtgtgaatt taaaagcatt tacatggcca
      721 agaaacctgt gaagctgcct ggctatcact atgtggatta caaactggac atcacctctc
      781 ataatgaaga ttatacggtc gtagagcaat atgagcgcgc cgaggctcgt catcatctgt
      841 ttcagtaaat gtccagacct gcaggcatgc aagctctaga ggcatcaaat aaaacgaaag
      901 gctcagtcga aagactgggc ctttcgtttt atctgttgaa atgcaccaaa aactcgtaaa
      961 agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc
     1021 tttgatatct aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
     1081 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
     1141 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat gcttactttg
     1201 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
     1261 gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct gatgtaatgg ttgttggtat
     1321 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
     1381 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
     1441 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
     1501 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
     1561 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
     1621 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
     1681 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
     1741 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
     1801 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
     1861 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
     1921 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
     1981 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
     2041 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
     2101 atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt
     2161 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
     2221 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
     2281 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
     2341 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
     2401 aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt
     2461 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
     2521 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
     2581 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
     2641 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
     2701 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
     2761 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtcaaccag acccgccagg
     2821 attctcatgc ataaaaaacg cccggcggca accgagcgtt ctgaattaat taatcatcgc
     2881 gactgcagag atcccagagt ccttattaga agaactcgtc aagaaggcga tagaaggcga
     2941 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     3001 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     3061 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     3121 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     3181 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     3241 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     3301 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     3361 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     3421 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     3481 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     3541 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     3601 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     3661 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     3721 tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc
     3781 agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccgacg
     3841 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     3901 ttcgtcttca c
//
