LOCUS       sequence-451741-        3909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the H-Ras membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3909)
  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 3909)
  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: 1_T7-mScarlet3-Hras
FEATURES             Location/Qualifiers
     source          1..3909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..73)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(70..92)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(256..844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(336..355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(961..985)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1085..1876,1877..1945))
                     /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     1708..1727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1946..2050)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2149..2171
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2163..2180
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2164..2180
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2233..2252
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2233..2251
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2319..2340
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             2385..3071
                     /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     2783..2802
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
       61 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      121 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      181 agaacatgtg agcaaaaggc cagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      841 tcaagaagat cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt
      901 cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg tcctcgtgat accaattcgg
      961 agcctgcttt tttgtacaaa cttgttgata atggcaattc aaggatcttc acctagatcc
     1021 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1081 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1141 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1201 gccccagtgc tgcaatgata ccgcgagagc cacgctcacc ggctccagat ttatcagcaa
     1261 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1321 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1381 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1441 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1501 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1561 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1621 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1681 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1741 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1801 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1861 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1921 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1981 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2041 gggttccgcg cacatttccc cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg
     2101 ccaaggaagt ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac
     2161 gttgtaaaac gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat
     2221 tccgcctgac cttaatacga ctcactatag ggcgaattgg gtaaagtgca agttaaagtg
     2281 aatcaattaa aagtaaccag caaccaagta aatcaactgc aactactgaa atctgccaag
     2341 aagtaattat tgaatacaag aagagaactc tgggcgccac aaccatggat agcaccgagg
     2401 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
     2461 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2521 tgagggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
     2581 tgtacggctc cagggccttc acgaagcacc ccgccgacat ccccgactac tggaagcagt
     2641 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgctgtgt
     2701 ccgtggccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg
     2761 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcat
     2821 ccaccgagcg gttgtacccc gaggacgtcg tgctgaaggg cgacattaag atggccctgc
     2881 gcctgaagga cggcggtcgc tacctggcgg acttcaagac cacctacagg gccaagaagc
     2941 ccgtgcagat gcccggcgct ttcaacatcg accgcaagtt ggacatcaca tcccacaacg
     3001 aggactacac cgtggtggaa cagtacgaac gctccgtggc ccgccactcc accggcggct
     3061 ccggtggctc ccgtgagatc cggcagcaca agctgcggaa gctgaaccct cctgatgaga
     3121 gtggccccgg ctgcatgagc tgcaagtgtg tgctctccta agcgatcgct atcgatgtta
     3181 taagttctga accgagaaag aggggagaat gtagagccga tagctgccca atactgccac
     3241 gcccccagca gcccaggcat ggaaaataat acagagaaaa tccacactta ttaggtgcta
     3301 ctgtaatttt ttttaaatcg cttaaaatta ttcggcgttt gcctagtgca aggcccattt
     3361 atggacaatt gtttgccaaa ctcaaagttt gccggtcgga ggaaaagtta caacgccaaa
     3421 atgtgttgaa aaatgcaaaa gaaattatta aaaacattaa aaaattatta aataatctat
     3481 gtaattcttg aatgtaaact ctgtgtaaat actaaaaatg aaaaacaaaa tggaaagcgt
     3541 agatggagaa cagttgaatc gttatatatt taaattaata ttgcatttat ttccaaatta
     3601 cagtgggaga agcgctgacc gacttcttgc agtgtactct cgaaataaaa cccctgtctg
     3661 taccacgtaa aacccctact aaaaattagt ctcgcccgct gctcacagtt agtttagttc
     3721 aattatacgc tactgtttaa tttattaact gtacttatta atgcgcgtca tgaataatga
     3781 ttgactgtcc cttatttttt ttttttgcta atttctgtat taaatttaat gatttaccat
     3841 aaaaatataa ataaaaatga cggtcgagtt gtaaaattaa taaaaaaaaa aaacgaatgc
     3901 tgcggccgc
//
