LOCUS       sequence-200517-        3929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding TurboRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3929)
  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 3929)
  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-TurboRFP
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..883
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      921..967
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1140..2090)
                     /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      2138..2360
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2866..2900)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2941..3735)
                     /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(3052..3071)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3662..3681
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(3914..3929,1..3)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggccctttcg tcttcaccac agctaacacc acgtcgtccc tatctgctgc cctaggtcta
       61 tgagtggttg ctggataact ttacgggcat gcataaggct cgtatgatat attcagggag
      121 accacaacgg tttccctcta caaataattt tgtttaactt ttctagattt aagaaggaga
      181 tatacatatg tctgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
      241 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
      301 ccagaccatg aagatcaagg tggtcgaggg cggcccttta cccttcgcct tcgacatcct
      361 ggctaccagc ttcatgtacg gcagcaaggc cttcatcaac cacacccagg gcatccccga
      421 cttctttaag cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga
      481 cgggggcgtg ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa
      541 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacaag
      601 gggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctga ggggccacag
      661 ccagatggcc ctgaagttag tgggcggggg ctacctgcac tgcagcttca agaccacata
      721 cagatccaag aaacccgcta agaacttgaa gatgcccggc ttccacttcg tggaccacag
      781 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt
      841 ggccaagtac tgcgacctgc ctagcaaact ggggcacagg taaatgtcca gacctgcagg
      901 catgcaagct ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      961 gttttatctg ttgaaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
     1021 caccgttttc atctgtgcat atggacagtt ttccctttga tatctaacgg tgaacagttg
     1081 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
     1141 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
     1201 catgagaacg aaccattgag atcatgctta ctttgcatgt cactcaaaaa ttttgcctca
     1261 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
     1321 cgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
     1381 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
     1441 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
     1501 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
     1561 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
     1621 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
     1681 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
     1741 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
     1801 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
     1861 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
     1921 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
     1981 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
     2041 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
     2101 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     2161 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     2221 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     2281 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     2341 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     2401 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     2461 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     2521 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     2581 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     2641 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     2701 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     2761 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     2821 gcttacccgt cttactgtca accagacccg ccaggattct catgcataaa aaacgcccgg
     2881 cggcaaccga gcgttctgaa ttaattaatc atcgcgactg cagagatccc agagtcctta
     2941 ttagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     3001 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3061 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     3121 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     3181 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     3241 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     3301 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     3361 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     3421 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     3481 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     3541 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     3601 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3661 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3721 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
     3781 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     3841 cttaccagag ggcgccccag ctggcaattc cgacgtctaa gaaaccatta ttatcatgac
     3901 attaacctat aaaaataggc gtatcacga
//
