LOCUS       sequence-448042-        3865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the Lyn membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3865)
  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 3865)
  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: 6_T7-Lyn-mScarlet3
FEATURES             Location/Qualifiers
     source          1..3865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..97)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      492..501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(594..615)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     complement(682..701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(683..701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(754..771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(754..770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(763..785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        884..988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(989..1057,1058..1849)
                     /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     complement(1207..1226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1949..1973
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      2090..2678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2579..2598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2831..2853
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2850..2866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2850..2866
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(join(3674..3865,1..495))
                     /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"
ORIGIN
        1 cagggccatc ttaatgtcgc ccttcagcac gacgtcctcg gggtacaacc gctcggtgga
       61 tgcttcccag cccattgtct tcttctgcat tacggggccg tcaggaggga agttggtgcc
      121 gcggagcttc accttgtaga tcagggtgcc gtcctccagg gaggtgtcct gggccacgga
      181 cacagcgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga
      241 ctgcttccag tagtcgggga tgtcggcggg gtgcttcgtg aaggccctgg agccgtacat
      301 gaactgaggg gacaggatgt cccaggagaa gggcaggggg ccacccttgg tcaccctcag
      361 cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc
      421 gtggccgttc atggagccct ccatgtgcac cttgaaccgc atgaactcct tgatcactgc
      481 ctcggtgcta tccatggcgg cctcgtcgtc gttcaggttg tccttgcgct tgctcttgat
      541 gcagcccatg gttgtggcgc ccagagttct cttcttgtat tcaataatta cttcttggca
      601 gatttcagta gttgcagttg atttacttgg ttgctggtta cttttaattg attcacttta
      661 acttgcactt tacccaattc gccctatagt gagtcgtatt aaggtcaggc ggaatggcac
      721 ttccttattc cagatgcgtg cggattatgc atgactggcc gtcgttttac aacgtcgtga
      781 ctgggaaaac cctggcgctt gtggtggatc acagttattg gagacttcct tggccgtacg
      841 atgggttttg tttcaggtat ctggcacttt tcggggaaat gtgcgcggaa cccctatttg
      901 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      961 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1021 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1081 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1141 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1201 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1261 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1321 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1381 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1441 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1501 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1561 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1621 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1681 taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg
     1741 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1801 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1861 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1921 ggtgaagatc cttgaattgc cattatcaac aagtttgtac aaaaaagcag gctccgaatt
     1981 ggtatcacga ggacgatttg aagcccccta cccatttacg ctgacttgac gggacggcgg
     2041 ctttgttgaa tagagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2221 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2701 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttccagcg acatatgcca
     2761 tacgccgaac tgaactccac ctaatgtctt ggagattgtc ggatagcgcg gaataggatt
     2821 gtgccctctg agcggataac aatttcacac aggaaacagc tatgaccatg acacatgcat
     2881 ccaccatcgc agacttatca tcactgagcc tccacctagc ctgcggccgc agcattcgtt
     2941 tttttttttt attaatttta caactcgacc gtcattttta tttatatttt tatggtaaat
     3001 cattaaattt aatacagaaa ttagcaaaaa aaaaaaataa gggacagtca atcattattc
     3061 atgacgcgca ttaataagta cagttaataa attaaacagt agcgtataat tgaactaaac
     3121 taactgtgag cagcgggcga gactaatttt tagtaggggt tttacgtggt acagacaggg
     3181 gttttatttc gagagtacac tgcaagaagt cggtcagcgc ttctcccact gtaatttgga
     3241 aataaatgca atattaattt aaatatataa cgattcaact gttctccatc tacgctttcc
     3301 attttgtttt tcatttttag tatttacaca gagtttacat tcaagaatta catagattat
     3361 ttaataattt tttaatgttt ttaataattt cttttgcatt tttcaacaca ttttggcgtt
     3421 gtaacttttc ctccgaccgg caaactttga gtttggcaaa caattgtcca taaatgggcc
     3481 ttgcactagg caaacgccga ataattttaa gcgatttaaa aaaaattaca gtagcaccta
     3541 ataagtgtgg attttctctg tattattttc catgcctggg ctgctggggg cgtggcagta
     3601 ttgggcagct atcggctcta cattctcccc tctttctcgg ttcagaactt ataacatcga
     3661 tagcgatcgc ttaggagcca ccggagccgc cggtggagtg gcgggccacg gagcgttcgt
     3721 actgttccac cacggtgtag tcctcgttgt gggatgtgat gtccaacttg cggtcgatgt
     3781 tgaaagcgcc gggcatctgc acgggcttct tggccctgta ggtggtcttg aagtccgcca
     3841 ggtagcgacc gccgtccttc aggcg
//
