LOCUS       sequence-275238-        7352 bp    DNA     circular SYN 15-APR-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 7352)
  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 7352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..586)
                     /label=P element 5' end
     CDS             join(898..900,901..903,904..1617)
                     /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(943..964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1204..1223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1551..1572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1733..1814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1770..1789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1824..1843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1861..1894
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             1906..1959
                     /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             1960..4605
                     /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(4614..4647)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(4720..4921,4922..4952)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(5058..5075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5229..5817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5309..5328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5988..6779,6780..6848))
                     /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     6611..6630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6849..6953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7116..7137)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 catgatgaaa taacataagg tggtcccgtc gatagccgaa gcttaccgaa gtatacactt
       61 aaattcagtg cacgtttgct tgttgagagg aaaggttgtg tgcggacgaa tttttttttg
      121 aaaacattaa cccttacgtg gaataaaaaa aaatgaaata ttgcaaattt tgctgcaaag
      181 ctgtgactgg agtaaaatta attcacgtgc cgaagtgtgc tattaagaga aaattgtggg
      241 agcagagcct tgggtgcagc cttggtgaaa actcccaaat ttgtgatacc cactttaatg
      301 attcgcagtg gaaggctgca cctgcaaaag gtcagacatt taaaaggagg cgactcaacg
      361 cagatgccgt acctagtaaa gtgatagagc ctgaaccaga aaagataaaa gaaggctata
      421 ccagtgggag tacacaaaca gagtaagttt gaatagtaaa aaaaatcatt tatgtaaaca
      481 ataacgtgac tgtgcgttag gtcctgttca ttgtttaatg aaaataagag cttgagggaa
      541 aaaattcgta ctttggagta cgaaatgcgt cgtttagagc agcagcggta ccgggccccc
      601 cctcgaggtc gacgaagttc ctattcttca aatagtatag gaacttcaag cttcttaagg
      661 aattcgagct cgcccgggga tctaattcaa ttagagacta attcaattag agctaattca
      721 attaggatca gcttatcgat ttcgaaccct cgaccgccgg agtataaata gaggcgcttc
      781 gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct
      841 aagcaaataa acaagcgcag ctgaacaagc taaacaatcg ggctagacct aggcaaaatg
      901 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1021 aagctgaccc tgaagctcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1081 gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     1141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1321 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1381 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1501 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1561 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaacca
     1621 tggctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1681 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cgaattcgta
     1861 cgcgcccggg gagcccaagg gcacgccctg gcacccggcg cgccagaggg cagaggcagc
     1921 ctgctcacct gcggcgacgt ggaagagaac cctgggccta tgaagctact gtcttctatc
     1981 gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga aaaaccgaag
     2041 tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac caaaaggtct
     2101 ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact ggaacagcta
     2161 tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga ttctttacag
     2221 gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa agatgccgtc
     2281 acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca gcatagaata
     2341 agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt gactgtatcg
     2401 attgactcgg cagctcatca tgataactcc acaattccgt tggattttat gcccagggat
     2461 gctcttcatg gatttgattg gtctgaagag gatgacatgt cggatggctt gcccttcctg
     2521 aaaacggacc ccaacaataa tgggttcttt ggcgacggtt ctctcttatg tattcttcga
     2581 tctattggct ttaaaccgga aaattacacg aactctaacg ttaacaggct cccgaccatg
     2641 attacggata gatacacgtt ggcttctaga tccacaacat cccgtttact tcaaagttat
     2701 ctcaataatt ttcaccccta ctgccctatc gtgcactcac cgacgctaat gatgttgtat
     2761 aataaccaga ttgaaatcgc gtcgaaggat caatggcaaa tcctttttaa ctgcatatta
     2821 gccattggag cctggtgtat agagggggaa tctactgata tagatgtttt ttactatcaa
     2881 aatgctaaat ctcatttgac gagcaaggtc ttcgagtcag gttccataat tttggtgaca
     2941 gccctacatc ttctgtcgcg atatacacag tggaggcaga aaacaaatac tagctataat
     3001 tttcacagct tttccataag aatggccata tcattgggct tgaataggga cctcccctcg
     3061 tccttcagtg atagcagcat tctggaacaa agacgccgaa tttggtggtc tgtctactct
     3121 tgggagatcc aattgtccct gctttatggt cgatccatcc agctttctca gaatacaatc
     3181 tccttccctt cttctgtcga cgatgtgcag cgtaccacaa caggtcccac catatatcat
     3241 ggcatcattg aaacagcaag gctcttacaa gttttcacaa aaatctatga actagacaaa
     3301 acagtaactg cagaaaaaag tcctatatgt gcaaaaaaat gcttgatgat ttgtaatgag
     3361 attgaggagg tttcgagaca ggcaccaaag tttttacaaa tggatatttc caccaccgct
     3421 ctaaccaatt tgttgaagga acacccttgg ctatccttta caagattcga actgaagtgg
     3481 aaacagttgt ctcttatcat ttatgtatta agagattttt tcactaattt tacccagaaa
     3541 aagtcacaac tagaacagga tcaaaatgat catcaaagtt atgaagttaa acgatgctcc
     3601 atcatgttaa gcgatgcagc acaaagaact gttatgtctg taagtagcta tatggacaat
     3661 cataatgtca ccccatattt tgcctggaat tgttcttatt acttgttcaa tgcagtccta
     3721 gtacccataa agactctact ctcaaactca aaatcgaatg ctgagaataa cgagaccgca
     3781 caattattac aacaaattaa cactgttctg atgctattaa aaaaactggc cacttttaaa
     3841 atccagactt gtgaaaaata cattcaagta ctggaagagg tatgtgcgcc gtttctgtta
     3901 tcacagtgtg caatcccatt accgcatatc agttataaca atagtaatgg tagcgccatt
     3961 aaaaatattg tcggttctgc aactatcgcc caatacccta ctcttccgga ggaaaatgtc
     4021 aacaatatca gtgttaaata tgtttctcct ggctcagtag ggccttcacc tgtgccattg
     4081 aaatcaggag caagtttcag tgatctagtc aagctgttat ctaaccgtcc accctctcgt
     4141 aactctccag tgacaatacc aagaagcaca ccttcgcatc gctcagtcac gccttttcta
     4201 gggcaacagc aacagctgca atcattagtg ccactgaccc cgtctgcttt gtttggtggc
     4261 gccaatttta atcaaagtgg gaatattgct gatagctcat tgtccttcac tttcactaac
     4321 agtagcaacg gtccgaacct cataacaact caaacaaatt ctcaagcgct ttcacaacca
     4381 attgcctcct ctaacgttca tgataacttc atgaataatg aaatcacggc tagtaaaatt
     4441 gatgatggta ataattcaaa accactgtca cctggttgga cggaccaaac tgcgtataac
     4501 gcgtttggaa tcactacagg gatgtttaat accactacaa tggatgatgt atataactat
     4561 ctattcgatg atgaagatac cccaccaaac ccaaaaaaag agtaagcggc cgcataactt
     4621 cgtatagcat acattatacg aagttatacc ggtggccgcg aagttcctat tcttcaaata
     4681 gtataggaac ttcgagctcc cggagagatg cagaagcttt gcgtactcgc aaattattaa
     4741 aaataaaact ttaaaaataa tttcgtctaa ttaatattat gagttaattc aaaccccacg
     4801 gacatgctaa gggttaatca acaatcatat cgctgtctca ctcagactca atacgacact
     4861 cagaatacta ttcctttcac tcgcacttat tgcaagcata cgttaagtgg atgtctcttg
     4921 ccgacgggac caccttatgt tatttcatca tggtctggcc attctcatcg tgagcttccg
     4981 ggtgctcgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5041 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5101 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5161 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5221 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5281 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5341 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5401 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5461 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5521 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5581 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5641 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5701 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5761 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5821 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5881 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5941 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6001 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6061 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6121 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6181 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6241 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6301 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6361 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6421 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6481 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6541 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6601 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6661 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6721 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6781 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6841 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6901 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6961 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     7021 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     7081 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     7141 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     7201 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     7261 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcc ttctccgccg
     7321 ccgatctgcc gctggactac gtgggtctgg cc
//
