LOCUS       sequence-275244-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal mScarlet-I fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  AUTHORS   Kondo S, Takahashi T, Yamagata N, Imanishi Y, Katow H, Hiramatsu S, 
            Lynn K, Abe A, Kumaraswamy A, Tanimoto H
  TITLE     Neurochemical Organization of the Drosophila Brain Visualized by 
            Endogenously Tagged Neurotransmitter Receptors.
  JOURNAL   Cell Rep. 2020 Jan 7;30(1):284-297.e5. doi: 
            10.1016/j.celrep.2019.12.018.
  PUBMED    31914394
REFERENCE   2  (bases 1 to 5354)
  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"; doi: 
            "10.1016/j.celrep.2019.12"; journalName: "Cell Rep"; date: 
            "2020-01-7- 7"; volume: "30"; issue: "1"; pages: "284-297.e5"
COMMENT     Sequence Label: pF3BGX-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    28..61
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     CDS             join(312..314,315..317,318..1031)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(357..378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(618..637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     965..986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1147..1228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1184..1203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1238..1257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1275..1308
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             1323..2018
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1456..1474)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1724..1743
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2030..2063)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2076..2109
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    join(2136..2337,2338..2368)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(2474..2491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2645..3233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2725..2744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3404..4195,4196..4264))
                     /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     4027..4046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4265..4369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4532..4553)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4769..4799,4800..5354)
                     /label=P element 5' end
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacgaa gttcctattc ttcaaatagt ataggaactt
       61 caagcttctt aaggaattcg agctcgcccg gggatctaat tcaattagag actaattcaa
      121 ttagagctaa ttcaattagg atcagcttat cgatttcgaa ccctcgaccg ccggagtata
      181 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
      241 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atcgggctag
      301 acctaggcaa aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      361 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      421 atgccaccta cggcaagctg accctgaagc tcatctgcac caccggcaag ctgcccgtgc
      481 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
      541 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      601 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      661 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      721 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
      781 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
      841 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      901 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
      961 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1021 tgtacaagta accatggcta gatcataatc agccatacca catttgtaga ggttttactt
     1081 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1141 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1261 gtatcgaatt cgtacgcgcc cggggagccc aagggcacgc cctggcaccc ggcgcgccca
     1321 aaatggtgag caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     1381 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1441 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     1501 acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca
     1561 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1621 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     1681 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     1741 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     1801 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     1861 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     1921 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     1981 gccgccactc caccggcggc atggacgagc tgtacaagta agcggccgca taacttcgta
     2041 tagcatacat tatacgaagt tataccggtg gccgcgaagt tcctattctt caaatagtat
     2101 aggaacttcg agctcccgga gagatgcaga agctttgcgt actcgcaaat tattaaaaat
     2161 aaaactttaa aaataatttc gtctaattaa tattatgagt taattcaaac cccacggaca
     2221 tgctaagggt taatcaacaa tcatatcgct gtctcactca gactcaatac gacactcaga
     2281 atactattcc tttcactcgc acttattgca agcatacgtt aagtggatgt ctcttgccga
     2341 cgggaccacc ttatgttatt tcatcatggt ctggccattc tcatcgtgag cttccgggtg
     2401 ctcgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4381 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4441 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4501 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4561 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4621 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4681 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggccttct ccgccgccga
     4741 tctgccgctg gactacgtgg gtctggccca tgatgaaata acataaggtg gtcccgtcga
     4801 tagccgaagc ttaccgaagt atacacttaa attcagtgca cgtttgcttg ttgagaggaa
     4861 aggttgtgtg cggacgaatt tttttttgaa aacattaacc cttacgtgga ataaaaaaaa
     4921 atgaaatatt gcaaattttg ctgcaaagct gtgactggag taaaattaat tcacgtgccg
     4981 aagtgtgcta ttaagagaaa attgtgggag cagagccttg ggtgcagcct tggtgaaaac
     5041 tcccaaattt gtgataccca ctttaatgat tcgcagtgga aggctgcacc tgcaaaaggt
     5101 cagacattta aaaggaggcg actcaacgca gatgccgtac ctagtaaagt gatagagcct
     5161 gaaccagaaa agataaaaga aggctatacc agtgggagta cacaaacaga gtaagtttga
     5221 atagtaaaaa aaatcattta tgtaaacaat aacgtgactg tgcgttaggt cctgttcatt
     5281 gtttaatgaa aataagagct tgagggaaaa aattcgtact ttggagtacg aaatgcgtcg
     5341 tttagagcag cagc
//
