LOCUS       sequence-275181-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RMCE vector for generating C-terminal Tango fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  AUTHORS   Katow H, Takahashi T, Saito K, Tanimoto H, Kondo S
  TITLE     Tango knock-ins visualize endogenous activity of G protein-coupled 
            receptors in Drosophila.
  JOURNAL   J Neurogenet. 2019 Mar - Jun;33(2):44-51. doi: 
            10.1080/01677063.2019.1611806. Epub 2019 May 14.
  PUBMED    31084242
REFERENCE   2  (bases 1 to 6095)
  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"; journalName: "J 
            Neurogenet. 2019 Mar - Jun"; volume: "33(2)"; pages: "44-51. doi: 
            10.1080/01677063.2019.1611806. Epub 2019 May 14"
COMMENT     Sequence Label: pF3BGX-TangoG-lexA
FEATURES             Location/Qualifiers
     source          1..6095
                     /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"
     primer_bind     complement(357..378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(618..637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     965..986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     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             1320..1340
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1341..1589,1590..1943)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="KALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEEH
                     LKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSLF
                     KPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGNK
                     VELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             2388..2759
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     protein_bind    complement(2771..2804)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2817..2850
                     /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(2877..3078,3079..3109)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(3215..3232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3386..3974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3466..3485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4145..4936,4937..5005))
                     /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     4768..4787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5006..5110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5273..5294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5510..5540,5541..6095)
                     /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 ggcgcgccag
     1321 agaacctgta cttccagggc aaggctctca cggcccgaca acaggaagtt tttgatttga
     1381 tacgggatca tatatcccaa acgggtatgc ctccgacccg cgcagagata gcacagcgac
     1441 tgggctttcg atcgcctaac gccgcggagg agcacttgaa ggcactggcc cgcaagggtg
     1501 tcattgaaat cgtgtccggt gcgagccgcg gaatccggct gttgcaggag gaggaagagg
     1561 gcctgccact ggtgggacgc gtggccgctg gcgagccgct gctggcccag caacacatag
     1621 agggacacta tcaggtggac ccctccttgt ttaagccaaa tgctgatttc ctgttgcggg
     1681 tgtcgggaat gtccatgaag gacatcggta ttatggatgg tgacctcctc gccgtccata
     1741 agacacagga tgtcaggaac ggccaggtag tcgttgccag gatagacgat gaggtcactg
     1801 tgaaacgtct caagaagcaa ggcaataagg tcgagctgct gccggagaat agcgagttca
     1861 agccgatcgt ggtggatctg cgacagcagt cctttactat cgagggcttg gccgtgggtg
     1921 tgatccgcaa cggagattgg ctgggatcca cgccgatgga gttccagtac ctgcccgata
     1981 cggatgaccg tcaccgtatc gaagaaaagc ggaagcgaac ctatgaaacc ttcaagtcca
     2041 tcatgaaaaa gtcccccttc tcgggcccca cggacccgcg ccccccgccc cgtcgtattg
     2101 cggttccttc gcgcagcagt gccagcgtcc ccaaacccgc accgcagccc tacccgttca
     2161 cttcctccct tagcacgatt aactatgatg agttccccac gatggtgttc cccagtggac
     2221 aaatttccca ggcatcggca ctggctccgg ccccaccgca agtcctcccc caggcgcccg
     2281 ctccggcacc ggctcccgca atggtgagtg ctctggccca ggcccccgct ccagtccccg
     2341 tgctggcgcc tggaccccca caggcagttg cccctcctgc tccgaaacca acgcaggcgg
     2401 gcgaaggaac cctgagcgag gccctcttgc agcttcagtt cgatgacgaa gacttgggag
     2461 ccctgctggg taacagcaca gaccctgccg tattcaccga tctcgcatcc gtggacaaca
     2521 gcgagtttca gcagctcttg aatcagggaa tcccggtcgc acctcatacc acagagccca
     2581 tgctgatgga atacccggag gctatcacgc gactggtgac cggcgcacag cgaccacccg
     2641 atccagcccc tgccccactg ggtgccccgg gtttgcccaa tggcctcctc agcggcgatg
     2701 aggatttctc cagcatcgct gatatggatt tctccgcttt gctgagccag ataagctcct
     2761 aagcggccgc ataacttcgt atagcataca ttatacgaag ttataccggt ggccgcgaag
     2821 ttcctattct tcaaatagta taggaacttc gagctcccgg agagatgcag aagctttgcg
     2881 tactcgcaaa ttattaaaaa taaaacttta aaaataattt cgtctaatta atattatgag
     2941 ttaattcaaa ccccacggac atgctaaggg ttaatcaaca atcatatcgc tgtctcactc
     3001 agactcaata cgacactcag aatactattc ctttcactcg cacttattgc aagcatacgt
     3061 taagtggatg tctcttgccg acgggaccac cttatgttat ttcatcatgg tctggccatt
     3121 ctcatcgtga gcttccgggt gctcgccagc tgcattaatg aatcggccaa cgcgcgggga
     3181 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3241 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3301 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3361 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3421 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3481 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3541 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3601 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3661 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3721 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3781 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3841 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3901 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3961 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4021 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4081 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4141 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4201 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4261 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4321 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4381 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4441 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4501 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4561 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4621 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4681 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4741 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4801 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4861 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4921 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4981 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5041 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5101 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     5161 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5221 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5281 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5341 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5461 gccggccttc tccgccgccg atctgccgct ggactacgtg ggtctggccc atgatgaaat
     5521 aacataaggt ggtcccgtcg atagccgaag cttaccgaag tatacactta aattcagtgc
     5581 acgtttgctt gttgagagga aaggttgtgt gcggacgaat ttttttttga aaacattaac
     5641 ccttacgtgg aataaaaaaa aatgaaatat tgcaaatttt gctgcaaagc tgtgactgga
     5701 gtaaaattaa ttcacgtgcc gaagtgtgct attaagagaa aattgtggga gcagagcctt
     5761 gggtgcagcc ttggtgaaaa ctcccaaatt tgtgataccc actttaatga ttcgcagtgg
     5821 aaggctgcac ctgcaaaagg tcagacattt aaaaggaggc gactcaacgc agatgccgta
     5881 cctagtaaag tgatagagcc tgaaccagaa aagataaaag aaggctatac cagtgggagt
     5941 acacaaacag agtaagtttg aatagtaaaa aaaatcattt atgtaaacaa taacgtgact
     6001 gtgcgttagg tcctgttcat tgtttaatga aaataagagc ttgagggaaa aaattcgtac
     6061 tttggagtac gaaatgcgtc gtttagagca gcagc
//
