LOCUS       sequence-275179-        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-TangoL-lexA
FEATURES             Location/Qualifiers
     source          1..6095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..586)
                     /label=P element 5' end
     protein_bind    614..647
                     /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(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             join(1927..2175,2176..2529)
                     /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             2974..3345
                     /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(3357..3390)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3403..3436
                     /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(3463..3664,3665..3695)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     complement(3801..3818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3972..4560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4052..4071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4731..5522,5523..5591))
                     /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     5354..5373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5592..5696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5859..5880)
                     /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 cgccagagaa cctgtacttc
     1921 cagctaaagg ctctcacggc ccgacaacag gaagtttttg atttgatacg ggatcatata
     1981 tcccaaacgg gtatgcctcc gacccgcgca gagatagcac agcgactggg ctttcgatcg
     2041 cctaacgccg cggaggagca cttgaaggca ctggcccgca agggtgtcat tgaaatcgtg
     2101 tccggtgcga gccgcggaat ccggctgttg caggaggagg aagagggcct gccactggtg
     2161 ggacgcgtgg ccgctggcga gccgctgctg gcccagcaac acatagaggg acactatcag
     2221 gtggacccct ccttgtttaa gccaaatgct gatttcctgt tgcgggtgtc gggaatgtcc
     2281 atgaaggaca tcggtattat ggatggtgac ctcctcgccg tccataagac acaggatgtc
     2341 aggaacggcc aggtagtcgt tgccaggata gacgatgagg tcactgtgaa acgtctcaag
     2401 aagcaaggca ataaggtcga gctgctgccg gagaatagcg agttcaagcc gatcgtggtg
     2461 gatctgcgac agcagtcctt tactatcgag ggcttggccg tgggtgtgat ccgcaacgga
     2521 gattggctgg gatccacgcc gatggagttc cagtacctgc ccgatacgga tgaccgtcac
     2581 cgtatcgaag aaaagcggaa gcgaacctat gaaaccttca agtccatcat gaaaaagtcc
     2641 cccttctcgg gccccacgga cccgcgcccc ccgccccgtc gtattgcggt tccttcgcgc
     2701 agcagtgcca gcgtccccaa acccgcaccg cagccctacc cgttcacttc ctcccttagc
     2761 acgattaact atgatgagtt ccccacgatg gtgttcccca gtggacaaat ttcccaggca
     2821 tcggcactgg ctccggcccc accgcaagtc ctcccccagg cgcccgctcc ggcaccggct
     2881 cccgcaatgg tgagtgctct ggcccaggcc cccgctccag tccccgtgct ggcgcctgga
     2941 cccccacagg cagttgcccc tcctgctccg aaaccaacgc aggcgggcga aggaaccctg
     3001 agcgaggccc tcttgcagct tcagttcgat gacgaagact tgggagccct gctgggtaac
     3061 agcacagacc ctgccgtatt caccgatctc gcatccgtgg acaacagcga gtttcagcag
     3121 ctcttgaatc agggaatccc ggtcgcacct cataccacag agcccatgct gatggaatac
     3181 ccggaggcta tcacgcgact ggtgaccggc gcacagcgac cacccgatcc agcccctgcc
     3241 ccactgggtg ccccgggttt gcccaatggc ctcctcagcg gcgatgagga tttctccagc
     3301 atcgctgata tggatttctc cgctttgctg agccagataa gctcctaagc ggccgcataa
     3361 cttcgtatag catacattat acgaagttat accggtggcc gcgaagttcc tattcttcaa
     3421 atagtatagg aacttcgagc tcccggagag atgcagaagc tttgcgtact cgcaaattat
     3481 taaaaataaa actttaaaaa taatttcgtc taattaatat tatgagttaa ttcaaacccc
     3541 acggacatgc taagggttaa tcaacaatca tatcgctgtc tcactcagac tcaatacgac
     3601 actcagaata ctattccttt cactcgcact tattgcaagc atacgttaag tggatgtctc
     3661 ttgccgacgg gaccacctta tgttatttca tcatggtctg gccattctca tcgtgagctt
     3721 ccgggtgctc gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3781 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3841 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3901 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4381 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4561 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5041 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5341 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5701 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5761 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5821 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5881 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5941 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6001 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gccttctccg
     6061 ccgccgatct gccgctggac tacgtgggtc tggcc
//
