LOCUS       sequence-448048-        4300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro synthesis of mRNA for fluorescent protein mScarlet3 tagged 
            with the LactC2 membrane localisation signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4300)
  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 4300)
  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: 9_T7-mScarlet3-LactC2
FEATURES             Location/Qualifiers
     source          1..4300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        59..77
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     145..166
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             211..897
                     /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     609..628
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2194..2210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2194..2210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2207..2229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2382..2970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2462..2481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3087..3111)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3211..4002,4003..4071))
                     /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     3834..3853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4072..4176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4275..4297
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4289..4300,1..6)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(4290..4300,1..6)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gccagtcatg cataatccgc acgcatctgg aataaggaag tgccattccg cctgacctta
       61 atacgactca ctatagggcg aattgggtaa agtgcaagtt aaagtgaatc aattaaaagt
      121 aaccagcaac caagtaaatc aactgcaact actgaaatct gccaagaagt aattattgaa
      181 tacaagaaga gaactctggg cgccacaacc atggatagca ccgaggcagt gatcaaggag
      241 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
      301 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgag ggtgaccaag
      361 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
      421 gccttcacga agcaccccgc cgacatcccc gactactgga agcagtcctt ccccgagggc
      481 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ctgtgtccgt ggcccaggac
      541 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
      601 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcatccac cgagcggttg
      661 taccccgagg acgtcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
      721 ggtcgctacc tggcggactt caagaccacc tacagggcca agaagcccgt gcagatgccc
      781 ggcgctttca acatcgaccg caagttggac atcacatccc acaacgagga ctacaccgtg
      841 gtggaacagt acgaacgctc cgtggcccgc cactccaccg gcggctccgg tggctcctcc
      901 ggactcagat cttgcactga acccctaggc ctgaaggata ataccatccc caacaagcag
      961 atcacagcct ccagctacta caaaacctgg ggcctgagtg cctttagctg gtttccctac
     1021 tacgcacgac tggataatca gggcaagttc aacgcctgga ccgcccagac caacagtgcc
     1081 tctgagtggc tgcagattga cctgggctcc cagaagcgag tcacgggcat catcacccag
     1141 ggtgcccgag actttggcca cattcaatat gtggctgcct acagggtggc ctatggtgat
     1201 gatggtgtga cctggactga gtacaaggac ccgggggcct cagaaagcaa gattttccct
     1261 ggtaacatgg acaataattc ccacaagaag aacatatttg agacgccgtt ccaggctcgc
     1321 ttcgtgcgga tccagcccgt ggcctggcac aaccgtatca ccctgcgagt ggagctgctg
     1381 ggctgttaag cgatcgctat cgatgttata agttctgaac cgagaaagag gggagaatgt
     1441 agagccgata gctgcccaat actgccacgc ccccagcagc ccaggcatgg aaaataatac
     1501 agagaaaatc cacacttatt aggtgctact gtaatttttt ttaaatcgct taaaattatt
     1561 cggcgtttgc ctagtgcaag gcccatttat ggacaattgt ttgccaaact caaagtttgc
     1621 cggtcggagg aaaagttaca acgccaaaat gtgttgaaaa atgcaaaaga aattattaaa
     1681 aacattaaaa aattattaaa taatctatgt aattcttgaa tgtaaactct gtgtaaatac
     1741 taaaaatgaa aaacaaaatg gaaagcgtag atggagaaca gttgaatcgt tatatattta
     1801 aattaatatt gcatttattt ccaaattaca gtgggagaag cgctgaccga cttcttgcag
     1861 tgtactctcg aaataaaacc cctgtctgta ccacgtaaaa cccctactaa aaattagtct
     1921 cgcccgctgc tcacagttag tttagttcaa ttatacgcta ctgtttaatt tattaactgt
     1981 acttattaat gcgcgtcatg aataatgatt gactgtccct tatttttttt ttttgctaat
     2041 ttctgtatta aatttaatga tttaccataa aaatataaat aaaaatgacg gtcgagttgt
     2101 aaaattaata aaaaaaaaaa acgaatgctg cggccgcagg ctaggtggag gctcagtgat
     2161 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     2221 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     2281 tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag
     2341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3001 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
     3061 aaatcgtcct cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg
     3121 caattcaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg
     3361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4081 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga
     4201 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     4261 agcgccaggg ttttcccagt cacgacgttg taaaacgacg
//
