LOCUS       sequence-448049-        4060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with a signal peptide and the CD8 transmembrane domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4060)
  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 4060)
  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: 10_T7-SP-CD8tm-mScarlet3
FEATURES             Location/Qualifiers
     source          1..4060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(136..240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     339..361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     353..370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     354..370
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     423..442
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        423..441
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     509..530
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             827..1510
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="DSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKL
                     RVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGAV
                     SVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKMA
                     LRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHST
                     GGSGGS"
     primer_bind     1222..1241
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2318..2334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2331..2353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2506..3094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2586..2605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3211..3235)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3335..4060,1..66,67..135))
                     /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     3958..3977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
       61 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
      181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
      241 cacatttccc cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg ccaaggaagt
      301 ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac
      361 gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac
      421 cttaatacga ctcactatag ggcgaattgg gtaaagtgca agttaaagtg aatcaattaa
      481 aagtaaccag caaccaagta aatcaactgc aactactgaa atctgccaag aagtaattat
      541 tgaatacaag aagagaactc tgggcgccac aaccatggat atcagctata tcttcgtcat
      601 ctgcctgatg gccctgtgca gcggcggcag cagtctcagc caagtcgagg gcaaacagaa
      661 atccggaagg ggccggggct ccatgtgggg caaggggacc ggattggact tcgcctgtga
      721 tatctacatc tgggcaccct tggccggcat ctgcgtggcc cttctgctgt ccttgatcat
      781 cactctcatc tgctaccaca gccgcacggg ccgcggtggt ggcggagata gcaccgaggc
      841 agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga
      901 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      961 gagggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat
     1021 gtacggctcc agggccttca cgaagcaccc cgccgacatc cccgactact ggaagcagtc
     1081 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgctgtgtc
     1141 cgtggcccag gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg
     1201 caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcatc
     1261 caccgagcgg ttgtaccccg aggacgtcgt gctgaagggc gacattaaga tggccctgcg
     1321 cctgaaggac ggcggtcgct acctggcgga cttcaagacc acctacaggg ccaagaagcc
     1381 cgtgcagatg cccggcgctt tcaacatcga ccgcaagttg gacatcacat cccacaacga
     1441 ggactacacc gtggtggaac agtacgaacg ctccgtggcc cgccactcca ccggcggctc
     1501 cggtggctcc taagcgatcg ctatcgatgt tataagttct gaaccgagaa agaggggaga
     1561 atgtagagcc gatagctgcc caatactgcc acgcccccag cagcccaggc atggaaaata
     1621 atacagagaa aatccacact tattaggtgc tactgtaatt ttttttaaat cgcttaaaat
     1681 tattcggcgt ttgcctagtg caaggcccat ttatggacaa ttgtttgcca aactcaaagt
     1741 ttgccggtcg gaggaaaagt tacaacgcca aaatgtgttg aaaaatgcaa aagaaattat
     1801 taaaaacatt aaaaaattat taaataatct atgtaattct tgaatgtaaa ctctgtgtaa
     1861 atactaaaaa tgaaaaacaa aatggaaagc gtagatggag aacagttgaa tcgttatata
     1921 tttaaattaa tattgcattt atttccaaat tacagtggga gaagcgctga ccgacttctt
     1981 gcagtgtact ctcgaaataa aacccctgtc tgtaccacgt aaaaccccta ctaaaaatta
     2041 gtctcgcccg ctgctcacag ttagtttagt tcaattatac gctactgttt aatttattaa
     2101 ctgtacttat taatgcgcgt catgaataat gattgactgt cccttatttt ttttttttgc
     2161 taatttctgt attaaattta atgatttacc ataaaaatat aaataaaaat gacggtcgag
     2221 ttgtaaaatt aataaaaaaa aaaaacgaat gctgcggccg caggctaggt ggaggctcag
     2281 tgatgataag tctgcgatgg tggatgcatg tgtcatggtc atagctgttt cctgtgtgaa
     2341 attgttatcc gctcagaggg cacaatccta ttccgcgcta tccgacaatc tccaagacat
     2401 taggtggagt tcagttcggc gtatggcata tgtcgctgga aagaacatgt gagcaaaagg
     2461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2941 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3121 gtctgacgct ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat gggtaggggg
     3181 cttcaaatcg tcctcgtgat accaattcgg agcctgcttt tttgtacaaa cttgttgata
     3241 atggcaattc aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc
     3481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4021 gaaaactctc aaggatctta ccgctgttga gatccagttc
//
