LOCUS       sequence-448046-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the PH membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4339)
  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 4339)
  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: 8_T7-PH-mScarlet3
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..95
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     87..104
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     88..104
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     157..176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     243..264
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             837..1523
                     /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     1235..1254
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2331..2347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2331..2347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2344..2366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2519..3107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2599..2618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3224..3248)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3348..4139,4140..4208))
                     /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     3971..3990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4209..4313)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag
       61 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca taatccgcac
      121 gcatctggaa taaggaagtg ccattccgcc tgaccttaat acgactcact atagggcgaa
      181 ttgggtaaag tgcaagttaa agtgaatcaa ttaaaagtaa ccagcaacca agtaaatcaa
      241 ctgcaactac tgaaatctgc caagaagtaa ttattgaata caagaagaga actctgggcg
      301 ccacaaccat ggactcgggc cgggacttcc tgaccctgca cggcctacag gatgatgagg
      361 atctacaggc gctgctgaag ggcagccagc tcctgaaggt gaagtccagc tcatggagga
      421 gagagcgctt ctacaagttg caggaggact gcaagaccat ctggcaggag tcccgcaagg
      481 tcatgcggac cccggagtcc cagctgttct ccatcgagga cattcaggag gtgcgaatgg
      541 ggcaccgcac ggagggtctg gagaagttcg cccgtgatgt gcccgaggac cgctgcttct
      601 ccattgtctt caaggaccag cgcaatacac tagacctcat cgccccatcg ccagctgatg
      661 cccagcactg ggtgctgggg ctgcacaaga tcatccacca ctcaggctcc atggaccagc
      721 gtcagaagct acagcactgg attcactcct gcttgcgaaa agctgacaaa aacaaggaca
      781 acaagatgag cttcaaggag ctgcagaact tcctgaagga gctcaacatc ctcgagatgg
      841 atagcaccga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca
      901 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
      961 agaccgccaa gctgagggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt
     1021 cccctcagtt catgtacggc tccagggcct tcacgaagca ccccgccgac atccccgact
     1081 actggaagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1141 gcggcgctgt gtccgtggcc caggacacct ccctggagga cggcaccctg atctacaagg
     1201 tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg
     1261 gctgggaagc atccaccgag cggttgtacc ccgaggacgt cgtgctgaag ggcgacatta
     1321 agatggccct gcgcctgaag gacggcggtc gctacctggc ggacttcaag accacctaca
     1381 gggccaagaa gcccgtgcag atgcccggcg ctttcaacat cgaccgcaag ttggacatca
     1441 catcccacaa cgaggactac accgtggtgg aacagtacga acgctccgtg gcccgccact
     1501 ccaccggcgg ctccggtggc tcctaagcga tcgctatcga tgttataagt tctgaaccga
     1561 gaaagagggg agaatgtaga gccgatagct gcccaatact gccacgcccc cagcagccca
     1621 ggcatggaaa ataatacaga gaaaatccac acttattagg tgctactgta atttttttta
     1681 aatcgcttaa aattattcgg cgtttgccta gtgcaaggcc catttatgga caattgtttg
     1741 ccaaactcaa agtttgccgg tcggaggaaa agttacaacg ccaaaatgtg ttgaaaaatg
     1801 caaaagaaat tattaaaaac attaaaaaat tattaaataa tctatgtaat tcttgaatgt
     1861 aaactctgtg taaatactaa aaatgaaaaa caaaatggaa agcgtagatg gagaacagtt
     1921 gaatcgttat atatttaaat taatattgca tttatttcca aattacagtg ggagaagcgc
     1981 tgaccgactt cttgcagtgt actctcgaaa taaaacccct gtctgtacca cgtaaaaccc
     2041 ctactaaaaa ttagtctcgc ccgctgctca cagttagttt agttcaatta tacgctactg
     2101 tttaatttat taactgtact tattaatgcg cgtcatgaat aatgattgac tgtcccttat
     2161 tttttttttt tgctaatttc tgtattaaat ttaatgattt accataaaaa tataaataaa
     2221 aatgacggtc gagttgtaaa attaataaaa aaaaaaaacg aatgctgcgg ccgcaggcta
     2281 ggtggaggct cagtgatgat aagtctgcga tggtggatgc atgtgtcatg gtcatagctg
     2341 tttcctgtgt gaaattgtta tccgctcaga gggcacaatc ctattccgcg ctatccgaca
     2401 atctccaaga cattaggtgg agttcagttc ggcgtatggc atatgtcgct ggaaagaaca
     2461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3121 tcttttctac ggggtctgac gctctattca acaaagccgc cgtcccgtca agtcagcgta
     3181 aatgggtagg gggcttcaaa tcgtcctcgt gataccaatt cggagcctgc ttttttgtac
     3241 aaacttgttg ataatggcaa ttcaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3481 ataccgcgag agccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4321 ccccgaaaag tgccagata
//
