LOCUS       sequence-275254-        5375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal Venus 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-Venus
FEATURES             Location/Qualifiers
     source          1..5375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(44..74,75..629)
                     /label=P element 5' end
     protein_bind    657..690
                     /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(941..943,944..946,947..1660)
                     /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    1776..1857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1813..1832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1867..1886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1904..1937
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             join(1952..1954,1955..1957,1958..2671)
                     /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"
     protein_bind    complement(2680..2713)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2726..2759
                     /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(2786..2987,2988..3018)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(3124..3141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3295..3883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3375..3394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4054..4845,4846..4914))
                     /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     4677..4696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4915..5019)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5182..5203)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttctccgcc gccgatctgc cgctggacta cgtgggtctg gcccatgatg aaataacata
       61 aggtggtccc gtcgatagcc gaagcttacc gaagtataca cttaaattca gtgcacgttt
      121 gcttgttgag aggaaaggtt gtgtgcggac gaattttttt ttgaaaacat taacccttac
      181 gtggaataaa aaaaaatgaa atattgcaaa ttttgctgca aagctgtgac tggagtaaaa
      241 ttaattcacg tgccgaagtg tgctattaag agaaaattgt gggagcagag ccttgggtgc
      301 agccttggtg aaaactccca aatttgtgat acccacttta atgattcgca gtggaaggct
      361 gcacctgcaa aaggtcagac atttaaaagg aggcgactca acgcagatgc cgtacctagt
      421 aaagtgatag agcctgaacc agaaaagata aaagaaggct ataccagtgg gagtacacaa
      481 acagagtaag tttgaatagt aaaaaaaatc atttatgtaa acaataacgt gactgtgcgt
      541 taggtcctgt tcattgttta atgaaaataa gagcttgagg gaaaaaattc gtactttgga
      601 gtacgaaatg cgtcgtttag agcagcagcg gtaccgggcc ccccctcgag gtcgacgaag
      661 ttcctattct tcaaatagta taggaacttc aagcttctta aggaattcga gctcgcccgg
      721 ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga tcagcttatc
      781 gatttcgaac cctcgaccgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat
      841 tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg
      901 cagctgaaca agctaaacaa tcgggctaga cctaggcaaa atggtgagca agggcgagga
      961 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1021 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     1081 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     1141 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1201 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1261 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1321 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1381 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1441 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     1501 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     1561 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1621 cgccgggatc actctcggca tggacgagct gtacaagtaa ccatggctag atcataatca
     1681 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1741 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1861 ctagttgtgg tttgtccaaa ctcatcaatg tatcgaattc gtacgcgccc ggggagccca
     1921 agggcacgcc ctggcacccg gcgcgcccaa aatggtgagc aagggcgagg agctgttcac
     1981 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2041 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tcatctgcac
     2101 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca
     2161 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2221 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2281 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2341 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2401 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     2461 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2521 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     2581 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2641 cactctcggc atggacgagc tgtacaagta agcggccgca taacttcgta tagcatacat
     2701 tatacgaagt tataccggtg gccgcgaagt tcctattctt caaatagtat aggaacttcg
     2761 agctcccgga gagatgcaga agctttgcgt actcgcaaat tattaaaaat aaaactttaa
     2821 aaataatttc gtctaattaa tattatgagt taattcaaac cccacggaca tgctaagggt
     2881 taatcaacaa tcatatcgct gtctcactca gactcaatac gacactcaga atactattcc
     2941 tttcactcgc acttattgca agcatacgtt aagtggatgt ctcttgccga cgggaccacc
     3001 ttatgttatt tcatcatggt ctggccattc tcatcgtgag cttccgggtg ctcgccagct
     3061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3481 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3721 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5041 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5101 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5161 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5221 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5281 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5341 agcccccgat ttagagcttg acggggaaag ccggc
//
