LOCUS       sequence-451742-        3877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the K-Ras 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: 2_T7-mScarlet3-KRas
FEATURES             Location/Qualifiers
     source          1..3877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     87..108
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             153..839
                     /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     551..570
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1713..1729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1713..1729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1726..1748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1901..2489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1981..2000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2606..2630)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2730..3521,3522..3590))
                     /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     3353..3372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3591..3695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3794..3816
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3808..3825
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3809..3825
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt aaagtgcaag ttaaagtgaa tcaattaaaa
       61 gtaaccagca accaagtaaa tcaactgcaa ctactgaaat ctgccaagaa gtaattattg
      121 aatacaagaa gagaactctg ggcgccacaa ccatggatag caccgaggca gtgatcaagg
      181 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
      241 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg agggtgacca
      301 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
      361 gggccttcac gaagcacccc gccgacatcc ccgactactg gaagcagtcc ttccccgagg
      421 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgctgtgtcc gtggcccagg
      481 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
      541 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcatcc accgagcggt
      601 tgtaccccga ggacgtcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
      661 gcggtcgcta cctggcggac ttcaagacca cctacagggc caagaagccc gtgcagatgc
      721 ccggcgcttt caacatcgac cgcaagttgg acatcacatc ccacaacgag gactacaccg
      781 tggtggaaca gtacgaacgc tccgtggccc gccactccac cggcggctcc ggtggctcca
      841 aagaaaagat gagcaaagat ggtaaaaaga agaaaaagaa gtcaaagaca aagtgtgtaa
      901 ttatgtaagc gatcgctatc gatgttataa gttctgaacc gagaaagagg ggagaatgta
      961 gagccgatag ctgcccaata ctgccacgcc cccagcagcc caggcatgga aaataataca
     1021 gagaaaatcc acacttatta ggtgctactg taattttttt taaatcgctt aaaattattc
     1081 ggcgtttgcc tagtgcaagg cccatttatg gacaattgtt tgccaaactc aaagtttgcc
     1141 ggtcggagga aaagttacaa cgccaaaatg tgttgaaaaa tgcaaaagaa attattaaaa
     1201 acattaaaaa attattaaat aatctatgta attcttgaat gtaaactctg tgtaaatact
     1261 aaaaatgaaa aacaaaatgg aaagcgtaga tggagaacag ttgaatcgtt atatatttaa
     1321 attaatattg catttatttc caaattacag tgggagaagc gctgaccgac ttcttgcagt
     1381 gtactctcga aataaaaccc ctgtctgtac cacgtaaaac ccctactaaa aattagtctc
     1441 gcccgctgct cacagttagt ttagttcaat tatacgctac tgtttaattt attaactgta
     1501 cttattaatg cgcgtcatga ataatgattg actgtccctt attttttttt tttgctaatt
     1561 tctgtattaa atttaatgat ttaccataaa aatataaata aaaatgacgg tcgagttgta
     1621 aaattaataa aaaaaaaaaa cgaatgctgc ggccgcaggc taggtggagg ctcagtgatg
     1681 ataagtctgc gatggtggat gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt
     1741 tatccgctca gagggcacaa tcctattccg cgctatccga caatctccaa gacattaggt
     1801 ggagttcagt tcggcgtatg gcatatgtcg ctggaaagaa catgtgagca aaaggccagc
     1861 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1921 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1981 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2041 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2101 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2161 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2221 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2281 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2341 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2401 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2461 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2521 acgctctatt caacaaagcc gccgtcccgt caagtcagcg taaatgggta gggggcttca
     2581 aatcgtcctc gtgataccaa ttcggagcct gcttttttgt acaaacttgt tgataatggc
     2641 aattcaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agagccacgc
     2881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccagat
     3721 acctgaaaca aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa
     3781 gcgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca
     3841 cgcatctgga ataaggaagt gccattccgc ctgacct
//
