LOCUS       sequence-267504-        7351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal T2A-GAL4 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7351)
  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 7351)
  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-T2A-GAL4
FEATURES             Location/Qualifiers
     source          1..7351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(44..74,75..628)
                     /label=P element 5' end
     protein_bind    656..689
                     /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(940..942,943..945,946..1659)
                     /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(985..1006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1246..1265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1593..1614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1775..1856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1812..1831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1866..1885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1903..1936
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             1948..2001
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2002..4647
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     protein_bind    complement(4656..4689)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4702..4735
                     /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(4762..4963,4964..4994)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(5100..5117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5271..5859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5351..5370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6030..6821,6822..6890))
                     /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     6653..6672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6891..6995)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7158..7179)
                     /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 gaaaaattcg tactttggag
      601 tacgaaatgc gtcgtttaga gcagcagcgg taccgggccc cccctcgagg tcgacgaagt
      661 tcctattctt caaatagtat aggaacttca agcttcttaa ggaattcgag ctcgcccggg
      721 gatctaattc aattagagac taattcaatt agagctaatt caattaggat cagcttatcg
      781 atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct tcgtctacgg agcgacaatt
      841 caattcaaac aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat aaacaagcgc
      901 agctgaacaa gctaaacaat cgggctagac ctaggcaaaa tggtgagcaa gggcgaggag
      961 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1021 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctc
     1081 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1141 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1201 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1261 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1321 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1381 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1441 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1501 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1561 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1621 gccgggatca ctctcggcat ggacgagctg tacaagtaac catggctaga tcataatcag
     1681 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1741 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1801 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1861 tagttgtggt ttgtccaaac tcatcaatgt atcgaattcg tacgcgcccg gggagcccaa
     1921 gggcacgccc tggcacccgg cgcgccagag ggcagaggca gcctgctcac ctgcggcgac
     1981 gtggaagaga accctgggcc tatgaagcta ctgtcttcta tcgaacaagc atgcgatatt
     2041 tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa gtgtctgaag
     2101 aacaactggg agtgtcgcta ctctcccaaa accaaaaggt ctccgctgac tagggcacat
     2161 ctgacagaag tggaatcaag gctagaaaga ctggaacagc tatttctact gatttttcct
     2221 cgagaagacc ttgacatgat tttgaaaatg gattctttac aggatataaa agcattgtta
     2281 acaggattat ttgtacaaga taatgtgaat aaagatgccg tcacagatag attggcttca
     2341 gtggagactg atatgcctct aacattgaga cagcatagaa taagtgcgac atcatcatcg
     2401 gaagagagta gtaacaaagg tcaaagacag ttgactgtat cgattgactc ggcagctcat
     2461 catgataact ccacaattcc gttggatttt atgcccaggg atgctcttca tggatttgat
     2521 tggtctgaag aggatgacat gtcggatggc ttgcccttcc tgaaaacgga ccccaacaat
     2581 aatgggttct ttggcgacgg ttctctctta tgtattcttc gatctattgg ctttaaaccg
     2641 gaaaattaca cgaactctaa cgttaacagg ctcccgacca tgattacgga tagatacacg
     2701 ttggcttcta gatccacaac atcccgttta cttcaaagtt atctcaataa ttttcacccc
     2761 tactgcccta tcgtgcactc accgacgcta atgatgttgt ataataacca gattgaaatc
     2821 gcgtcgaagg atcaatggca aatccttttt aactgcatat tagccattgg agcctggtgt
     2881 atagaggggg aatctactga tatagatgtt ttttactatc aaaatgctaa atctcatttg
     2941 acgagcaagg tcttcgagtc aggttccata attttggtga cagccctaca tcttctgtcg
     3001 cgatatacac agtggaggca gaaaacaaat actagctata attttcacag cttttccata
     3061 agaatggcca tatcattggg cttgaatagg gacctcccct cgtccttcag tgatagcagc
     3121 attctggaac aaagacgccg aatttggtgg tctgtctact cttgggagat ccaattgtcc
     3181 ctgctttatg gtcgatccat ccagctttct cagaatacaa tctccttccc ttcttctgtc
     3241 gacgatgtgc agcgtaccac aacaggtccc accatatatc atggcatcat tgaaacagca
     3301 aggctcttac aagttttcac aaaaatctat gaactagaca aaacagtaac tgcagaaaaa
     3361 agtcctatat gtgcaaaaaa atgcttgatg atttgtaatg agattgagga ggtttcgaga
     3421 caggcaccaa agtttttaca aatggatatt tccaccaccg ctctaaccaa tttgttgaag
     3481 gaacaccctt ggctatcctt tacaagattc gaactgaagt ggaaacagtt gtctcttatc
     3541 atttatgtat taagagattt tttcactaat tttacccaga aaaagtcaca actagaacag
     3601 gatcaaaatg atcatcaaag ttatgaagtt aaacgatgct ccatcatgtt aagcgatgca
     3661 gcacaaagaa ctgttatgtc tgtaagtagc tatatggaca atcataatgt caccccatat
     3721 tttgcctgga attgttctta ttacttgttc aatgcagtcc tagtacccat aaagactcta
     3781 ctctcaaact caaaatcgaa tgctgagaat aacgagaccg cacaattatt acaacaaatt
     3841 aacactgttc tgatgctatt aaaaaaactg gccactttta aaatccagac ttgtgaaaaa
     3901 tacattcaag tactggaaga ggtatgtgcg ccgtttctgt tatcacagtg tgcaatccca
     3961 ttaccgcata tcagttataa caatagtaat ggtagcgcca ttaaaaatat tgtcggttct
     4021 gcaactatcg cccaataccc tactcttccg gaggaaaatg tcaacaatat cagtgttaaa
     4081 tatgtttctc ctggctcagt agggccttca cctgtgccat tgaaatcagg agcaagtttc
     4141 agtgatctag tcaagctgtt atctaaccgt ccaccctctc gtaactctcc agtgacaata
     4201 ccaagaagca caccttcgca tcgctcagtc acgccttttc tagggcaaca gcaacagctg
     4261 caatcattag tgccactgac cccgtctgct ttgtttggtg gcgccaattt taatcaaagt
     4321 gggaatattg ctgatagctc attgtccttc actttcacta acagtagcaa cggtccgaac
     4381 ctcataacaa ctcaaacaaa ttctcaagcg ctttcacaac caattgcctc ctctaacgtt
     4441 catgataact tcatgaataa tgaaatcacg gctagtaaaa ttgatgatgg taataattca
     4501 aaaccactgt cacctggttg gacggaccaa actgcgtata acgcgtttgg aatcactaca
     4561 gggatgttta ataccactac aatggatgat gtatataact atctattcga tgatgaagat
     4621 accccaccaa acccaaaaaa agagtaagcg gccgcataac ttcgtatagc atacattata
     4681 cgaagttata ccggtggccg cgaagttcct attcttcaaa tagtatagga acttcgagct
     4741 cccggagaga tgcagaagct ttgcgtactc gcaaattatt aaaaataaaa ctttaaaaat
     4801 aatttcgtct aattaatatt atgagttaat tcaaacccca cggacatgct aagggttaat
     4861 caacaatcat atcgctgtct cactcagact caatacgaca ctcagaatac tattcctttc
     4921 actcgcactt attgcaagca tacgttaagt ggatgtctct tgccgacggg accaccttat
     4981 gttatttcat catggtctgg ccattctcat cgtgagcttc cgggtgctcg ccagctgcat
     5041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5701 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7021 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     7081 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     7141 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     7201 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     7261 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     7321 cccgatttag agcttgacgg ggaaagccgg c
//
