LOCUS       sequence-275249-        5375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal mTurqouise2 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5375)
  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 5375)
  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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5375
                     /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"
     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             join(1323..1325,1326..1328,1329..2042)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(2051..2084)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2097..2130
                     /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(2157..2358,2359..2389)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(2495..2512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2666..3254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2746..2765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3425..4216,4217..4285))
                     /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     4048..4067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4286..4390)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4553..4574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4790..4820,4821..5375)
                     /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 gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1381 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1441 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1501 ccctcgtgac caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga
     1561 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1621 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1681 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1741 acaagctgga gtacaactac tttagcgaca acgtctatat caccgccgac aagcagaaga
     1801 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     1861 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1921 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1981 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2041 aagcggccgc ataacttcgt atagcataca ttatacgaag ttataccggt ggccgcgaag
     2101 ttcctattct tcaaatagta taggaacttc gagctcccgg agagatgcag aagctttgcg
     2161 tactcgcaaa ttattaaaaa taaaacttta aaaataattt cgtctaatta atattatgag
     2221 ttaattcaaa ccccacggac atgctaaggg ttaatcaaca atcatatcgc tgtctcactc
     2281 agactcaata cgacactcag aatactattc ctttcactcg cacttattgc aagcatacgt
     2341 taagtggatg tctcttgccg acgggaccac cttatgttat ttcatcatgg tctggccatt
     2401 ctcatcgtga gcttccgggt gctcgccagc tgcattaatg aatcggccaa cgcgcgggga
     2461 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4381 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     4441 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4501 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4561 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4621 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4681 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4741 gccggccttc tccgccgccg atctgccgct ggactacgtg ggtctggccc atgatgaaat
     4801 aacataaggt ggtcccgtcg atagccgaag cttaccgaag tatacactta aattcagtgc
     4861 acgtttgctt gttgagagga aaggttgtgt gcggacgaat ttttttttga aaacattaac
     4921 ccttacgtgg aataaaaaaa aatgaaatat tgcaaatttt gctgcaaagc tgtgactgga
     4981 gtaaaattaa ttcacgtgcc gaagtgtgct attaagagaa aattgtggga gcagagcctt
     5041 gggtgcagcc ttggtgaaaa ctcccaaatt tgtgataccc actttaatga ttcgcagtgg
     5101 aaggctgcac ctgcaaaagg tcagacattt aaaaggaggc gactcaacgc agatgccgta
     5161 cctagtaaag tgatagagcc tgaaccagaa aagataaaag aaggctatac cagtgggagt
     5221 acacaaacag agtaagtttg aatagtaaaa aaaatcattt atgtaaacaa taacgtgact
     5281 gtgcgttagg tcctgttcat tgtttaatga aaataagagc ttgagggaaa aaattcgtac
     5341 tttggagtac gaaatgcgtc gtttagagca gcagc
//
