LOCUS       sequence-461232-        4063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with a signal peptide and a GPI attachment site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4063)
  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 4063)
  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: 11_T7-SP-mScarlet3-GPI
FEATURES             Location/Qualifiers
     source          1..4063
                     /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(245..833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(325..344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(950..974)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1074..1865,1866..1934))
                     /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     1697..1716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1935..2039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2138..2160
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2152..2169
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2153..2169
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2222..2241
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2222..2240
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2308..2329
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      2485..2494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2491..3177
                     /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     2889..2908
                     /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 ccaggaaccg taaaaaggcc gcgttgctgg
      241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      841 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
      901 agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga gcctgctttt
      961 ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct tttaaattaa
     1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1201 gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2041 acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc caaggaagtc
     2101 tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg
     2161 acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc
     2221 ttaatacgac tcactatagg gcgaattggg taaagtgcaa gttaaagtga atcaattaaa
     2281 agtaaccagc aaccaagtaa atcaactgca actactgaaa tctgccaaga agtaattatt
     2341 gaatacaaga agagaactct gggcgccaca accatggagc tcttttggag tatagtcttt
     2401 actgtcctcc tgagtttctc ctgccggggg tcagactggg aatctctgca gtcgacggta
     2461 ccgcgggccc gggatccacc ggtcgccacc atggatagca ccgaggcagt gatcaaggag
     2521 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     2581 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgag ggtgaccaag
     2641 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
     2701 gccttcacga agcaccccgc cgacatcccc gactactgga agcagtcctt ccccgagggc
     2761 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ctgtgtccgt ggcccaggac
     2821 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     2881 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcatccac cgagcggttg
     2941 taccccgagg acgtcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
     3001 ggtcgctacc tggcggactt caagaccacc tacagggcca agaagcccgt gcagatgccc
     3061 ggcgctttca acatcgaccg caagttggac atcacatccc acaacgagga ctacaccgtg
     3121 gtggaacagt acgaacgctc cgtggcccgc cactccaccg gcggctccgg tggctcctat
     3181 ggaggaaacg gctccggaca gcaccagtat gatccgcgcc caagcagcgg tcattctaga
     3241 cacagatatg cacttatacc cataccatta gcagtaatta caacatgtat tgtgctgtat
     3301 atgaatgttc tttaagcgat cgctatcgat gttataagtt ctgaaccgag aaagagggga
     3361 gaatgtagag ccgatagctg cccaatactg ccacgccccc agcagcccag gcatggaaaa
     3421 taatacagag aaaatccaca cttattaggt gctactgtaa ttttttttaa atcgcttaaa
     3481 attattcggc gtttgcctag tgcaaggccc atttatggac aattgtttgc caaactcaaa
     3541 gtttgccggt cggaggaaaa gttacaacgc caaaatgtgt tgaaaaatgc aaaagaaatt
     3601 attaaaaaca ttaaaaaatt attaaataat ctatgtaatt cttgaatgta aactctgtgt
     3661 aaatactaaa aatgaaaaac aaaatggaaa gcgtagatgg agaacagttg aatcgttata
     3721 tatttaaatt aatattgcat ttatttccaa attacagtgg gagaagcgct gaccgacttc
     3781 ttgcagtgta ctctcgaaat aaaacccctg tctgtaccac gtaaaacccc tactaaaaat
     3841 tagtctcgcc cgctgctcac agttagttta gttcaattat acgctactgt ttaatttatt
     3901 aactgtactt attaatgcgc gtcatgaata atgattgact gtcccttatt tttttttttt
     3961 gctaatttct gtattaaatt taatgattta ccataaaaat ataaataaaa atgacggtcg
     4021 agttgtaaaa ttaataaaaa aaaaaaacga atgctgcggc cgc
//
