LOCUS       sequence-275176-        6182 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 6182)
  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 6182)
  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-TangoVG-lexA
FEATURES             Location/Qualifiers
     source          1..6182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(127..129,130..132,133..846)
                     /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(172..193)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(433..452)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     780..801
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    962..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(999..1018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1053..1072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1090..1123
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             1222..1242
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1243..1491,1492..1845)
                     /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             2290..2661
                     /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(2673..2706)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2719..2752
                     /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(2779..2980,2981..3011)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(3117..3134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3288..3876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3368..3387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4047..4838,4839..4907))
                     /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     4670..4689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4908..5012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5175..5196)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5412..5442,5443..5997)
                     /label=P element 5' end
     protein_bind    6025..6058
                     /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)"
ORIGIN
        1 aggcgcttcg tctacggagc gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa
       61 gcgaaagcta agcaaataaa caagcgcagc tgaacaagct aaacaatcgg gctagaccta
      121 ggcaaaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      181 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      241 acctacggca agctgaccct gaagctcatc tgcaccaccg gcaagctgcc cgtgccctgg
      301 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
      361 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      421 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      481 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      541 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
      601 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      661 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      721 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
      781 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      841 aagtaaccat ggctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
      901 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
      961 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1021 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1081 gaattcgtac gcgcccgggg agcccaaggg cacgccctgg cacccggcgc gccaggacgc
     1141 accccaccca gcctgggtcc ccaagatgag tcctgcacca ccgccagctc ctccctggcc
     1201 aaggacactt catcgaccgg tgagaacctg tacttccagg gcaaggctct cacggcccga
     1261 caacaggaag tttttgattt gatacgggat catatatccc aaacgggtat gcctccgacc
     1321 cgcgcagaga tagcacagcg actgggcttt cgatcgccta acgccgcgga ggagcacttg
     1381 aaggcactgg cccgcaaggg tgtcattgaa atcgtgtccg gtgcgagccg cggaatccgg
     1441 ctgttgcagg aggaggaaga gggcctgcca ctggtgggac gcgtggccgc tggcgagccg
     1501 ctgctggccc agcaacacat agagggacac tatcaggtgg acccctcctt gtttaagcca
     1561 aatgctgatt tcctgttgcg ggtgtcggga atgtccatga aggacatcgg tattatggat
     1621 ggtgacctcc tcgccgtcca taagacacag gatgtcagga acggccaggt agtcgttgcc
     1681 aggatagacg atgaggtcac tgtgaaacgt ctcaagaagc aaggcaataa ggtcgagctg
     1741 ctgccggaga atagcgagtt caagccgatc gtggtggatc tgcgacagca gtcctttact
     1801 atcgagggct tggccgtggg tgtgatccgc aacggagatt ggctgggatc cacgccgatg
     1861 gagttccagt acctgcccga tacggatgac cgtcaccgta tcgaagaaaa gcggaagcga
     1921 acctatgaaa ccttcaagtc catcatgaaa aagtccccct tctcgggccc cacggacccg
     1981 cgccccccgc cccgtcgtat tgcggttcct tcgcgcagca gtgccagcgt ccccaaaccc
     2041 gcaccgcagc cctacccgtt cacttcctcc cttagcacga ttaactatga tgagttcccc
     2101 acgatggtgt tccccagtgg acaaatttcc caggcatcgg cactggctcc ggccccaccg
     2161 caagtcctcc cccaggcgcc cgctccggca ccggctcccg caatggtgag tgctctggcc
     2221 caggcccccg ctccagtccc cgtgctggcg cctggacccc cacaggcagt tgcccctcct
     2281 gctccgaaac caacgcaggc gggcgaagga accctgagcg aggccctctt gcagcttcag
     2341 ttcgatgacg aagacttggg agccctgctg ggtaacagca cagaccctgc cgtattcacc
     2401 gatctcgcat ccgtggacaa cagcgagttt cagcagctct tgaatcaggg aatcccggtc
     2461 gcacctcata ccacagagcc catgctgatg gaatacccgg aggctatcac gcgactggtg
     2521 accggcgcac agcgaccacc cgatccagcc cctgccccac tgggtgcccc gggtttgccc
     2581 aatggcctcc tcagcggcga tgaggatttc tccagcatcg ctgatatgga tttctccgct
     2641 ttgctgagcc agataagctc ctaagcggcc gcataacttc gtatagcata cattatacga
     2701 agttataccg gtggccgcga agttcctatt cttcaaatag tataggaact tcgagctccc
     2761 ggagagatgc agaagctttg cgtactcgca aattattaaa aataaaactt taaaaataat
     2821 ttcgtctaat taatattatg agttaattca aaccccacgg acatgctaag ggttaatcaa
     2881 caatcatatc gctgtctcac tcagactcaa tacgacactc agaatactat tcctttcact
     2941 cgcacttatt gcaagcatac gttaagtgga tgtctcttgc cgacgggacc accttatgtt
     3001 atttcatcat ggtctggcca ttctcatcgt gagcttccgg gtgctcgcca gctgcattaa
     3061 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3721 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3961 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4021 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4081 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4141 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4201 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4261 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4321 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4381 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4441 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4501 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4561 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4621 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4681 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4741 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4801 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4861 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4921 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4981 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5041 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5101 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5161 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5221 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5281 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5341 gatttagagc ttgacgggga aagccggcct tctccgccgc cgatctgccg ctggactacg
     5401 tgggtctggc ccatgatgaa ataacataag gtggtcccgt cgatagccga agcttaccga
     5461 agtatacact taaattcagt gcacgtttgc ttgttgagag gaaaggttgt gtgcggacga
     5521 attttttttt gaaaacatta acccttacgt ggaataaaaa aaaatgaaat attgcaaatt
     5581 ttgctgcaaa gctgtgactg gagtaaaatt aattcacgtg ccgaagtgtg ctattaagag
     5641 aaaattgtgg gagcagagcc ttgggtgcag ccttggtgaa aactcccaaa tttgtgatac
     5701 ccactttaat gattcgcagt ggaaggctgc acctgcaaaa ggtcagacat ttaaaaggag
     5761 gcgactcaac gcagatgccg tacctagtaa agtgatagag cctgaaccag aaaagataaa
     5821 agaaggctat accagtggga gtacacaaac agagtaagtt tgaatagtaa aaaaaatcat
     5881 ttatgtaaac aataacgtga ctgtgcgtta ggtcctgttc attgtttaat gaaaataaga
     5941 gcttgaggga aaaaattcgt actttggagt acgaaatgcg tcgtttagag cagcagcggt
     6001 accgggcccc ccctcgaggt cgacgaagtt cctattcttc aaatagtata ggaacttcaa
     6061 gcttcttaag gaattcgagc tcgcccgggg atctaattca attagagact aattcaatta
     6121 gagctaattc aattaggatc agcttatcga tttcgaaccc tcgaccgccg gagtataaat
     6181 ag
//
