LOCUS       sequence-275175-        6179 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 6179)
  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 6179)
  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-TangoVL-lexA
FEATURES             Location/Qualifiers
     source          1..6179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(213..243,244..445)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    complement(472..505)
                     /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)"
     protein_bind    518..551
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(563..934)
                     /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"
     CDS             complement(join(1379..1732,1733..1981))
                     /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"
     protein_bind    2098..2131
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     complement(2149..2168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2178..2259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2203..2222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2375..3088,3089..3091,3092..3094))
                     /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(2420..2441)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2769..2788
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3028..3049
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3345..3378)
                     /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(3406..3960,3961..3991)
                     /label=P element 5' end
     primer_bind     4207..4228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4391..4495
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4496..4564,4565..5356)
                     /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     complement(4714..4733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5527..6115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6016..6035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
       61 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      121 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcgag
      181 cacccggaag ctcacgatga gaatggccag accatgatga aataacataa ggtggtcccg
      241 tcggcaagag acatccactt aacgtatgct tgcaataagt gcgagtgaaa ggaatagtat
      301 tctgagtgtc gtattgagtc tgagtgagac agcgatatga ttgttgatta acccttagca
      361 tgtccgtggg gtttgaatta actcataata ttaattagac gaaattattt ttaaagtttt
      421 atttttaata atttgcgagt acgcaaagct tctgcatctc tccgggagct cgaagttcct
      481 atactatttg aagaatagga acttcgcggc caccggtata acttcgtata atgtatgcta
      541 tacgaagtta tgcggccgct taggagctta tctggctcag caaagcggag aaatccatat
      601 cagcgatgct ggagaaatcc tcatcgccgc tgaggaggcc attgggcaaa cccggggcac
      661 ccagtggggc aggggctgga tcgggtggtc gctgtgcgcc ggtcaccagt cgcgtgatag
      721 cctccgggta ttccatcagc atgggctctg tggtatgagg tgcgaccggg attccctgat
      781 tcaagagctg ctgaaactcg ctgttgtcca cggatgcgag atcggtgaat acggcagggt
      841 ctgtgctgtt acccagcagg gctcccaagt cttcgtcatc gaactgaagc tgcaagaggg
      901 cctcgctcag ggttccttcg cccgcctgcg ttggtttcgg agcaggaggg gcaactgcct
      961 gtgggggtcc aggcgccagc acggggactg gagcgggggc ctgggccaga gcactcacca
     1021 ttgcgggagc cggtgccgga gcgggcgcct gggggaggac ttgcggtggg gccggagcca
     1081 gtgccgatgc ctgggaaatt tgtccactgg ggaacaccat cgtggggaac tcatcatagt
     1141 taatcgtgct aagggaggaa gtgaacgggt agggctgcgg tgcgggtttg gggacgctgg
     1201 cactgctgcg cgaaggaacc gcaatacgac ggggcggggg gcgcgggtcc gtggggcccg
     1261 agaaggggga ctttttcatg atggacttga aggtttcata ggttcgcttc cgcttttctt
     1321 cgatacggtg acggtcatcc gtatcgggca ggtactggaa ctccatcggc gtggatccca
     1381 gccaatctcc gttgcggatc acacccacgg ccaagccctc gatagtaaag gactgctgtc
     1441 gcagatccac cacgatcggc ttgaactcgc tattctccgg cagcagctcg accttattgc
     1501 cttgcttctt gagacgtttc acagtgacct catcgtctat cctggcaacg actacctggc
     1561 cgttcctgac atcctgtgtc ttatggacgg cgaggaggtc accatccata ataccgatgt
     1621 ccttcatgga cattcccgac acccgcaaca ggaaatcagc atttggctta aacaaggagg
     1681 ggtccacctg atagtgtccc tctatgtgtt gctgggccag cagcggctcg ccagcggcca
     1741 cgcgtcccac cagtggcagg ccctcttcct cctcctgcaa cagccggatt ccgcggctcg
     1801 caccggacac gatttcaatg acacccttgc gggccagtgc cttcaagtgc tcctccgcgg
     1861 cgttaggcga tcgaaagccc agtcgctgtg ctatctctgc gcgggtcgga ggcatacccg
     1921 tttgggatat atgatcccgt atcaaatcaa aaacttcctg ttgtcgggcc gtgagagcct
     1981 ttagctggaa gtacaggttc tcaccggtcg atgaagtgtc cttggccagg gaggagctgg
     2041 cggtggtgca ggactcatct tggggaccca ggctgggtgg gggtcctggc gcgccgggtg
     2101 ccagggcgtg cccttgggct ccccgggcgc gtacgaattc gatacattga tgagtttgga
     2161 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2221 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2281 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2341 aaatgtggta tggctgatta tgatctagcc atggttactt gtacagctcg tccatgccga
     2401 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     2461 ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc
     2521 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga
     2581 tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat
     2641 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     2701 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     2761 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     2821 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
     2881 agcactgcag gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     2941 cggtggtgca gatgagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     3001 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     3061 ccccggtgaa cagctcctcg cccttgctca ccattttgcc taggtctagc ccgattgttt
     3121 agcttgttca gctgcgcttg tttatttgct tagctttcgc ttagcgacgt gttcactttg
     3181 cttgtttgaa ttgaattgtc gctccgtaga cgaagcgcct ctatttatac tccggcggtc
     3241 gagggttcga aatcgataag ctgatcctaa ttgaattagc tctaattgaa ttagtctcta
     3301 attgaattag atccccgggc gagctcgaat tccttaagaa gcttgaagtt cctatactat
     3361 ttgaagaata ggaacttcgt cgacctcgag ggggggcccg gtaccgctgc tgctctaaac
     3421 gacgcatttc gtactccaaa gtacgaattt tttccctcaa gctcttattt tcattaaaca
     3481 atgaacagga cctaacgcac agtcacgtta ttgtttacat aaatgatttt ttttactatt
     3541 caaacttact ctgtttgtgt actcccactg gtatagcctt cttttatctt ttctggttca
     3601 ggctctatca ctttactagg tacggcatct gcgttgagtc gcctcctttt aaatgtctga
     3661 ccttttgcag gtgcagcctt ccactgcgaa tcattaaagt gggtatcaca aatttgggag
     3721 ttttcaccaa ggctgcaccc aaggctctgc tcccacaatt ttctcttaat agcacacttc
     3781 ggcacgtgaa ttaattttac tccagtcaca gctttgcagc aaaatttgca atatttcatt
     3841 tttttttatt ccacgtaagg gttaatgttt tcaaaaaaaa attcgtccgc acacaacctt
     3901 tcctctcaac aagcaaacgt gcactgaatt taagtgtata cttcggtaag cttcggctat
     3961 cgacgggacc accttatgtt atttcatcat gggccagacc cacgtagtcc agcggcagat
     4021 cggcggcgga gaaggccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4321 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4381 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4441 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4681 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4801 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5041 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5101 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5161 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5221 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5341 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5401 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5461 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5521 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5581 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5641 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5701 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5761 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5821 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5881 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5941 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6001 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6061 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6121 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttct
//
