LOCUS       sequence-448025-        3877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the K-Ras6R membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3877)
  AUTHORS   Karapidaki I, Handberg-Thorsager M, Momose T, Yasuo H, Genikhovich 
            G, Assaf S, Deleau C, Pang Y, Clayton P, Lohmann B, Rusciano ML, 
            Stranges M, Mathieu J, Zilliox M, Ustyantsev K, Salmon B, 
            Laplace-Builhe B, Koenig M, Colgren JJ, Arnone MI, Berezikov E, 
            Brunet T, Bucher G, Burkhardt P, Dickinson DJ, Houliston E, Huisken 
            J, Leclere L, Averof M
  TITLE     A toolkit for testing membrane-localising tags across species
  JOURNAL   bioRxiv 2024.11.12.623055
REFERENCE   2  (bases 1 to 3877)
  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"; journalName: "bioRxiv 
            2024.11.12.623055"
COMMENT     Sequence Label: 3_T7-mScarlet3-KRas6R
FEATURES             Location/Qualifiers
     source          1..3877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1126..1142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1126..1142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1139..1161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1314..1902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1394..1413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2019..2043)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2143..2934,2935..3003))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2766..2785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3004..3108)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3207..3229
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3221..3238
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3222..3238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3291..3310
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3291..3309
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3377..3398
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             join(3443..3877,1..252)
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     3841..3860
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 tccaccgagc ggttgtaccc cgaggacgtc gtgctgaagg gcgacattaa gatggccctg
       61 cgcctgaagg acggcggtcg ctacctggcg gacttcaaga ccacctacag ggccaagaag
      121 cccgtgcaga tgcccggcgc tttcaacatc gaccgcaagt tggacatcac atcccacaac
      181 gaggactaca ccgtggtgga acagtacgaa cgctccgtgg cccgccactc caccggcggc
      241 tccggtggct ccaaagaaaa gatgagcaaa gatggtagaa ggagacgacg cagatcaaag
      301 acaaagtgtg taattatgta agcgatcgct atcgatgtta taagttctga accgagaaag
      361 aggggagaat gtagagccga tagctgccca atactgccac gcccccagca gcccaggcat
      421 ggaaaataat acagagaaaa tccacactta ttaggtgcta ctgtaatttt ttttaaatcg
      481 cttaaaatta ttcggcgttt gcctagtgca aggcccattt atggacaatt gtttgccaaa
      541 ctcaaagttt gccggtcgga ggaaaagtta caacgccaaa atgtgttgaa aaatgcaaaa
      601 gaaattatta aaaacattaa aaaattatta aataatctat gtaattcttg aatgtaaact
      661 ctgtgtaaat actaaaaatg aaaaacaaaa tggaaagcgt agatggagaa cagttgaatc
      721 gttatatatt taaattaata ttgcatttat ttccaaatta cagtgggaga agcgctgacc
      781 gacttcttgc agtgtactct cgaaataaaa cccctgtctg taccacgtaa aacccctact
      841 aaaaattagt ctcgcccgct gctcacagtt agtttagttc aattatacgc tactgtttaa
      901 tttattaact gtacttatta atgcgcgtca tgaataatga ttgactgtcc cttatttttt
      961 ttttttgcta atttctgtat taaatttaat gatttaccat aaaaatataa ataaaaatga
     1021 cggtcgagtt gtaaaattaa taaaaaaaaa aaacgaatgc tgcggccgca ggctaggtgg
     1081 aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc
     1141 tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc cgacaatctc
     1201 caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa gaacatgtga
     1261 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1321 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1381 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1441 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1501 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1561 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1621 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1681 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1741 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1801 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1861 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1921 tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg
     1981 gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt tgtacaaact
     2041 tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2281 gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3121 aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct ccaataactg
     3181 tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtca
     3241 tgcataatcc gcacgcatct ggaataagga agtgccattc cgcctgacct taatacgact
     3301 cactataggg cgaattgggt aaagtgcaag ttaaagtgaa tcaattaaaa gtaaccagca
     3361 accaagtaaa tcaactgcaa ctactgaaat ctgccaagaa gtaattattg aatacaagaa
     3421 gagaactctg ggcgccacaa ccatggatag caccgaggca gtgatcaagg agttcatgcg
     3481 gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg
     3541 cgagggccgc ccctacgagg gcacccagac cgccaagctg agggtgacca agggtggccc
     3601 cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca gggccttcac
     3661 gaagcacccc gccgacatcc ccgactactg gaagcagtcc ttccccgagg gcttcaagtg
     3721 ggagcgcgtg atgaacttcg aggacggcgg cgctgtgtcc gtggcccagg acacctccct
     3781 ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc ctcctgacgg
     3841 ccccgtaatg cagaagaaga caatgggctg ggaagca
//
