LOCUS       sequence-448045-        4072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the Src64B membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4072)
  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 4072)
  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: 7_T7-Src64B-mScarlet3
FEATURES             Location/Qualifiers
     source          1..4072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     143..160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     144..160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     213..232
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        213..231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     299..320
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             626..1312
                     /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     1024..1043
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2120..2136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2120..2136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2133..2155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2308..2896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2388..2407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3013..3037)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3137..3928,3929..3997))
                     /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     3760..3779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3998..4072,1..30))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cagatacctg
       61 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
      121 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
      181 ctggaataag gaagtgccat tccgcctgac cttaatacga ctcactatag ggcgaattgg
      241 gtaaagtgca agttaaagtg aatcaattaa aagtaaccag caaccaagta aatcaactgc
      301 aactactgaa atctgccaag aagtaattat tgaatacaag aagagaactc tgggcgccac
      361 aaccatgggc aacaaatgct gcagcaagcg acaggatcag gaactggcac tggcctatcc
      421 cactgggggc tacaagaaat ccgactacac ctttggccag acgcacatca acagcagcgg
      481 cggcggcaac atgggcggcg ttcttggcca gaagcataac aacggtggct cgctggactc
      541 gcgctacacg cccgatccca atcatcgggg tccgttgaaa atcggcggaa agggcggcgt
      601 tgacatcatc agaccacgcg gatccatgga tagcaccgag gcagtgatca aggagttcat
      661 gcggttcaag gtgcacatgg agggctccat gaacggccac gagttcgaga tcgagggcga
      721 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgagggtga ccaagggtgg
      781 ccccctgccc ttctcctggg acatcctgtc ccctcagttc atgtacggct ccagggcctt
      841 cacgaagcac cccgccgaca tccccgacta ctggaagcag tccttccccg agggcttcaa
      901 gtgggagcgc gtgatgaact tcgaggacgg cggcgctgtg tccgtggccc aggacacctc
      961 cctggaggac ggcaccctga tctacaaggt gaagctccgc ggcaccaact tccctcctga
     1021 cggccccgta atgcagaaga agacaatggg ctgggaagca tccaccgagc ggttgtaccc
     1081 cgaggacgtc gtgctgaagg gcgacattaa gatggccctg cgcctgaagg acggcggtcg
     1141 ctacctggcg gacttcaaga ccacctacag ggccaagaag cccgtgcaga tgcccggcgc
     1201 tttcaacatc gaccgcaagt tggacatcac atcccacaac gaggactaca ccgtggtgga
     1261 acagtacgaa cgctccgtgg cccgccactc caccggcggc tccggtggct cctaagcgat
     1321 cgctatcgat gttataagtt ctgaaccgag aaagagggga gaatgtagag ccgatagctg
     1381 cccaatactg ccacgccccc agcagcccag gcatggaaaa taatacagag aaaatccaca
     1441 cttattaggt gctactgtaa ttttttttaa atcgcttaaa attattcggc gtttgcctag
     1501 tgcaaggccc atttatggac aattgtttgc caaactcaaa gtttgccggt cggaggaaaa
     1561 gttacaacgc caaaatgtgt tgaaaaatgc aaaagaaatt attaaaaaca ttaaaaaatt
     1621 attaaataat ctatgtaatt cttgaatgta aactctgtgt aaatactaaa aatgaaaaac
     1681 aaaatggaaa gcgtagatgg agaacagttg aatcgttata tatttaaatt aatattgcat
     1741 ttatttccaa attacagtgg gagaagcgct gaccgacttc ttgcagtgta ctctcgaaat
     1801 aaaacccctg tctgtaccac gtaaaacccc tactaaaaat tagtctcgcc cgctgctcac
     1861 agttagttta gttcaattat acgctactgt ttaatttatt aactgtactt attaatgcgc
     1921 gtcatgaata atgattgact gtcccttatt tttttttttt gctaatttct gtattaaatt
     1981 taatgattta ccataaaaat ataaataaaa atgacggtcg agttgtaaaa ttaataaaaa
     2041 aaaaaaacga atgctgcggc cgcaggctag gtggaggctc agtgatgata agtctgcgat
     2101 ggtggatgca tgtgtcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcagag
     2161 ggcacaatcc tattccgcgc tatccgacaa tctccaagac attaggtgga gttcagttcg
     2221 gcgtatggca tatgtcgctg gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctctattcaa
     2941 caaagccgcc gtcccgtcaa gtcagcgtaa atgggtaggg ggcttcaaat cgtcctcgtg
     3001 ataccaattc ggagcctgct tttttgtaca aacttgttga taatggcaat tcaaggatct
     3061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3241 gcttaccatc tggccccagt gctgcaatga taccgcgaga gccacgctca ccggctccag
     3301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4021 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt at
//
