LOCUS       sequence-200534-        3931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding 2ndVal mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3931)
  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 3931)
  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 mRFP1
FEATURES             Location/Qualifiers
     source          1..3931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             197..871
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIK
                     MRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STG"
     terminator      933..979
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1152..2102)
                     /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      2150..2372
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2877..2911)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2952..3746)
                     /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(3063..3082)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3673..3692
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(3925..3931,1..12)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtatcacgag gccctttcgt cttcaccaca gctaacacca cgtcgtccct atctgctgcc
       61 ctaggtctat gactggttgc tggataactt tacgggcatg cataaggctc gtatgatata
      121 ttcagggaga ccacaacggt ttccctctac aaataatttt gtttaacttt tctagattta
      181 agaaggagat atacatatgg tggccagcag cgaggacgtg attaaagagt tcatgcgctt
      241 caaagttcgc atggagggtt ctgttaacgg tcacgagttc gagatcgaag gcgaaggcga
      301 gggccgtccg tatgaaggca cccagaccgc caaactgaaa gtgactaaag gcggcccgct
      361 gccttttgcg tgggacatcc tgagcccgca atttcagtac ggttctaaag cgtatgttaa
      421 acacccagcg gatatcccgg actatctgaa gctgtctttt ccggaaggtt tcaagtggga
      481 acgcgtaatg aattttgaag atggtggtgt cgtgaccgtc actcaggact cctccctgca
      541 ggatggcgag ttcatctata aagttaaact gcgtggtact aattttccat ctgatggccc
      601 ggtgatgcag aaaaagacga tgggttggga ggcgtctacc gaacgcatgt atccggaaga
      661 tggtgcgctg aaaggcgaaa ttaaaatgcg cctgaaactg aaagatggcg gccattatga
      721 cgctgaagtg aaaaccacgt acatggccaa gaaacctgtg cagctgcctg gcgcgtacaa
      781 gaccgatatt aaactggaca tcacctctca taatgaagat tatacgatcg tagagcaata
      841 tgagcgcgcg gagggtcgtc attctaccgg tggcatggat gaactataca aataaatgtc
      901 cagacctgca ggcatgcaag ctctagaggc atcaaataaa acgaaaggct cagtcgaaag
      961 actgggcctt tcgttttatc tgttgaaatg caccaaaaac tcgtaaaagc tctgatgtat
     1021 ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatctaac
     1081 ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag
     1141 ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt
     1201 ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat gtcactcaaa
     1261 aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt
     1321 cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc
     1381 atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact
     1441 tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg
     1501 taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca
     1561 tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt
     1621 gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag
     1681 tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg
     1741 aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg
     1801 gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca
     1861 gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt ttccctactg
     1921 atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc tttccttgta
     1981 gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt ggtttcatgc
     2041 tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac taattcagac
     2101 atacatctca attggtctag gtgattttaa tcactatacc aattgagatg ggctagtcaa
     2161 tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg ctagaccttt
     2221 gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt
     2281 ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata aaaaaagata
     2341 aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc gcagtattac
     2401 aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc
     2461 ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat
     2521 caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca cggctctggc
     2581 agtgaatggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     2641 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     2701 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     2761 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     2821 atcatcaaca ggcttacccg tcttactgtc aaccagaccc gccaggattc tcatgcataa
     2881 aaaacgcccg gcggcaaccg agcgttctga attaattaat catcgcgact gcagagatcc
     2941 cagagtcctt attagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3001 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3061 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     3121 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     3181 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     3241 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     3301 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3361 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3421 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3481 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     3541 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     3601 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     3661 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     3721 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     3781 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     3841 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtcta agaaaccatt
     3901 attatcatga cattaaccta taaaaatagg c
//
