LOCUS       sequence-451743-        4021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the H-RitC membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4021)
  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 4021)
  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: 4_T7-mScarlet3-RitC
FEATURES             Location/Qualifiers
     source          1..4021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     87..108
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             153..839
                     /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     551..570
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1857..1873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1857..1873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1870..1892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2045..2633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2125..2144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2750..2774)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(2874..3665,3666..3734))
                     /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     3497..3516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3735..3839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3938..3960
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3952..3969
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3953..3969
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt aaagtgcaag ttaaagtgaa tcaattaaaa
       61 gtaaccagca accaagtaaa tcaactgcaa ctactgaaat ctgccaagaa gtaattattg
      121 aatacaagaa gagaactctg ggcgccacaa ccatggatag caccgaggca gtgatcaagg
      181 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
      241 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg agggtgacca
      301 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
      361 gggccttcac gaagcacccc gccgacatcc ccgactactg gaagcagtcc ttccccgagg
      421 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgctgtgtcc gtggcccagg
      481 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
      541 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcatcc accgagcggt
      601 tgtaccccga ggacgtcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
      661 gcggtcgcta cctggcggac ttcaagacca cctacagggc caagaagccc gtgcagatgc
      721 ccggcgcttt caacatcgac cgcaagttgg acatcacatc ccacaacgag gactacaccg
      781 tggtggaaca gtacgaacgc tccgtggccc gccactccac cggcggctcc ggtggctccg
      841 tcgaccacaa gaaaaagtca aagtgtccct tttttgagac atctgctgca taccgctact
      901 atattgatga tgttttccat gcccttgtac gggagatacg taggaaagaa aaggaggcag
      961 tactggccat ggagaaaaaa tctaagccca aaaacagtgt atggaagagg ctaaaatcac
     1021 cattccggaa gaagaaagat tcagtaactt aagcgatcgc tatcgatgtt ataagttctg
     1081 aaccgagaaa gaggggagaa tgtagagccg atagctgccc aatactgcca cgcccccagc
     1141 agcccaggca tggaaaataa tacagagaaa atccacactt attaggtgct actgtaattt
     1201 tttttaaatc gcttaaaatt attcggcgtt tgcctagtgc aaggcccatt tatggacaat
     1261 tgtttgccaa actcaaagtt tgccggtcgg aggaaaagtt acaacgccaa aatgtgttga
     1321 aaaatgcaaa agaaattatt aaaaacatta aaaaattatt aaataatcta tgtaattctt
     1381 gaatgtaaac tctgtgtaaa tactaaaaat gaaaaacaaa atggaaagcg tagatggaga
     1441 acagttgaat cgttatatat ttaaattaat attgcattta tttccaaatt acagtgggag
     1501 aagcgctgac cgacttcttg cagtgtactc tcgaaataaa acccctgtct gtaccacgta
     1561 aaacccctac taaaaattag tctcgcccgc tgctcacagt tagtttagtt caattatacg
     1621 ctactgttta atttattaac tgtacttatt aatgcgcgtc atgaataatg attgactgtc
     1681 ccttattttt tttttttgct aatttctgta ttaaatttaa tgatttacca taaaaatata
     1741 aataaaaatg acggtcgagt tgtaaaatta ataaaaaaaa aaaacgaatg ctgcggccgc
     1801 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
     1861 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
     1921 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
     1981 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2041 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2101 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2161 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2221 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2281 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2341 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2401 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2461 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2521 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2581 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2641 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
     2701 agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga gcctgctttt
     2761 ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct tttaaattaa
     2821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2941 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3001 gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3181 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3481 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3721 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3841 acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc caaggaagtc
     3901 tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg
     3961 acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc
     4021 t
//
