LOCUS       sequence-448029-        3871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the GAP43 membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  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 3871)
  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: 5_T7-GAP43-mScarlet3
FEATURES             Location/Qualifiers
     source          1..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(230..334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     433..455
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     447..464
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     448..464
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     517..536
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        517..535
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     603..624
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             729..1415
                     /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     1127..1146
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2223..2239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2223..2239)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2236..2258)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2411..2999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2491..2510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3116..3140)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3240..3871,1..160,161..229))
                     /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     join(3863..3871,1..11)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
       61 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
      241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccagata
      361 cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag
      421 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca taatccgcac
      481 gcatctggaa taaggaagtg ccattccgcc tgaccttaat acgactcact atagggcgaa
      541 ttgggtaaag tgcaagttaa agtgaatcaa ttaaaagtaa ccagcaacca agtaaatcaa
      601 ctgcaactac tgaaatctgc caagaagtaa ttattgaata caagaagaga actctgggcg
      661 ccacaaccat gctgtgctgt atgagaagaa ccaaacaggt tgaaaagaat gatgaggacc
      721 aaaagatcat ggatagcacc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
      781 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      841 acgagggcac ccagaccgcc aagctgaggg tgaccaaggg tggccccctg cccttctcct
      901 gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcacgaag caccccgccg
      961 acatccccga ctactggaag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     1021 acttcgagga cggcggcgct gtgtccgtgg cccaggacac ctccctggag gacggcaccc
     1081 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     1141 agaagacaat gggctgggaa gcatccaccg agcggttgta ccccgaggac gtcgtgctga
     1201 agggcgacat taagatggcc ctgcgcctga aggacggcgg tcgctacctg gcggacttca
     1261 agaccaccta cagggccaag aagcccgtgc agatgcccgg cgctttcaac atcgaccgca
     1321 agttggacat cacatcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     1381 tggcccgcca ctccaccggc ggctccggtg gctcctaagc gatcgctatc gatgttataa
     1441 gttctgaacc gagaaagagg ggagaatgta gagccgatag ctgcccaata ctgccacgcc
     1501 cccagcagcc caggcatgga aaataataca gagaaaatcc acacttatta ggtgctactg
     1561 taattttttt taaatcgctt aaaattattc ggcgtttgcc tagtgcaagg cccatttatg
     1621 gacaattgtt tgccaaactc aaagtttgcc ggtcggagga aaagttacaa cgccaaaatg
     1681 tgttgaaaaa tgcaaaagaa attattaaaa acattaaaaa attattaaat aatctatgta
     1741 attcttgaat gtaaactctg tgtaaatact aaaaatgaaa aacaaaatgg aaagcgtaga
     1801 tggagaacag ttgaatcgtt atatatttaa attaatattg catttatttc caaattacag
     1861 tgggagaagc gctgaccgac ttcttgcagt gtactctcga aataaaaccc ctgtctgtac
     1921 cacgtaaaac ccctactaaa aattagtctc gcccgctgct cacagttagt ttagttcaat
     1981 tatacgctac tgtttaattt attaactgta cttattaatg cgcgtcatga ataatgattg
     2041 actgtccctt attttttttt tttgctaatt tctgtattaa atttaatgat ttaccataaa
     2101 aatataaata aaaatgacgg tcgagttgta aaattaataa aaaaaaaaaa cgaatgctgc
     2161 ggccgcaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca
     2221 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg
     2281 cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg
     2341 ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2821 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3001 aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt
     3061 caagtcagcg taaatgggta gggggcttca aatcgtcctc gtgataccaa ttcggagcct
     3121 gcttttttgt acaaacttgt tgataatggc aattcaagga tcttcaccta gatcctttta
     3181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3361 agtgctgcaa tgataccgcg agagccacgc tcaccggctc cagatttatc agcaataaac
     3421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3841 tcttgcccgg cgtcaatacg ggataatacc g
//
