LOCUS       sequence-275169-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal fusion of seven tandem GFP11 
            tags (split GFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  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 5126)
  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-7xGFP11
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(77..79,80..82,83..796)
                     /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(122..143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(383..402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     730..751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    912..993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(949..968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1003..1022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1040..1073
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             1130..1555
                     /codon_start=1
                     /product="seven tandem copies of the 11th beta-strand of 
                     superfolder GFP"
                     /label=GFP11x7
                     /note="each GFP11 associates with GFP(1-10), consisting of 
                     the first 10 beta-strands, to reconstitute fluorescent GFP 
                     (Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDH
                     MVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGITGGSG
                     GRDHMVLHEYVNAAGITGGSGGRDHMVLHEYVNAAGIT"
     protein_bind    complement(1567..1600)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1613..1646
                     /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(1673..1874,1875..1905)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(2011..2028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2182..2770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2262..2281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2941..3732,3733..3801))
                     /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     3564..3583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3802..3906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4069..4090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4306..4336,4337..4891)
                     /label=P element 5' end
     protein_bind    4919..4952
                     /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)"
ORIGIN
        1 acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatcgg
       61 gctagaccta ggcaaaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      121 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      181 gggcgatgcc acctacggca agctgaccct gaagctcatc tgcaccaccg gcaagctgcc
      241 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta
      301 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      361 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      421 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      481 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
      541 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
      601 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      661 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
      721 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      781 cgagctgtac aagtaaccat ggctagatca taatcagcca taccacattt gtagaggttt
      841 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
      901 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
      961 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1021 tcaatgtatc gaattcgtac gcgcccgggg agcccaaggg cacgccctgg cacccggcgc
     1081 gccaggtggc tctggaagtt caggtggagg ctcgggtggc ggcagttcgc gtgaccacat
     1141 ggtccttcat gagtatgtaa atgctgctgg gattacaggt ggctctggag gtagagatca
     1201 tatggttctc cacgaatacg ttaacgccgc aggcatcact ggcggtagtg gaggacgcga
     1261 ccatatggta ctacatgaat atgtcaatgc agccggaata accggagggt ccggaggccg
     1321 ggatcacatg gtgctgcatg agtatgtgaa cgcggcgggt ataactggtg ggtcgggcgg
     1381 acgagaccat atggtgcttc acgaatacgt aaacgcagct ggcattactg gcggatcagg
     1441 tggcagggat cacatggtac tccatgagta cgtgaacgct gctggaatca caggcggtag
     1501 cggcggtcgg gaccatatgg tcctgcacga atatgtcaat gctgccggta tcacctaagc
     1561 ggccgcataa cttcgtatag catacattat acgaagttat accggtggcc gcgaagttcc
     1621 tattcttcaa atagtatagg aacttcgagc tcccggagag atgcagaagc tttgcgtact
     1681 cgcaaattat taaaaataaa actttaaaaa taatttcgtc taattaatat tatgagttaa
     1741 ttcaaacccc acggacatgc taagggttaa tcaacaatca tatcgctgtc tcactcagac
     1801 tcaatacgac actcagaata ctattccttt cactcgcact tattgcaagc atacgttaag
     1861 tggatgtctc ttgccgacgg gaccacctta tgttatttca tcatggtctg gccattctca
     1921 tcgtgagctt ccgggtgctc gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1981 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2041 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2101 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2161 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2221 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2581 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3241 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3541 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3901 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     3961 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4021 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4081 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4141 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4201 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4261 gccttctccg ccgccgatct gccgctggac tacgtgggtc tggcccatga tgaaataaca
     4321 taaggtggtc ccgtcgatag ccgaagctta ccgaagtata cacttaaatt cagtgcacgt
     4381 ttgcttgttg agaggaaagg ttgtgtgcgg acgaattttt ttttgaaaac attaaccctt
     4441 acgtggaata aaaaaaaatg aaatattgca aattttgctg caaagctgtg actggagtaa
     4501 aattaattca cgtgccgaag tgtgctatta agagaaaatt gtgggagcag agccttgggt
     4561 gcagccttgg tgaaaactcc caaatttgtg atacccactt taatgattcg cagtggaagg
     4621 ctgcacctgc aaaaggtcag acatttaaaa ggaggcgact caacgcagat gccgtaccta
     4681 gtaaagtgat agagcctgaa ccagaaaaga taaaagaagg ctataccagt gggagtacac
     4741 aaacagagta agtttgaata gtaaaaaaaa tcatttatgt aaacaataac gtgactgtgc
     4801 gttaggtcct gttcattgtt taatgaaaat aagagcttga gggaaaaaat tcgtactttg
     4861 gagtacgaaa tgcgtcgttt agagcagcag cggtaccggg ccccccctcg aggtcgacga
     4921 agttcctatt cttcaaatag tataggaact tcaagcttct taaggaattc gagctcgccc
     4981 ggggatctaa ttcaattaga gactaattca attagagcta attcaattag gatcagctta
     5041 tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta cggagcgaca
     5101 attcaattca aacaagcaaa gtgaac
//
