LOCUS       sequence-381472-        5263 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent reporter for Gal4-induced Cre recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5263)
  AUTHORS   Yoshihara M, Nishino T, Sambe N, Nayakama T, Radtke F, Mizuno S, 
            Takahashi S
  TITLE     Generation of a Gal4-dependent gene recombination and illuminating 
            mouse.
  JOURNAL   Exp Anim. 2022 Aug 5;71(3):385-390. doi: 10.1538/expanim.21-0202. 
            Epub 2022 Apr 21.
  PUBMED    35444103
REFERENCE   2  (bases 1 to 5263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1538/expanim.21-0202"; journalName: "Exp Anim"; date: 
            "2022-08-5- 5"; volume: "71"; issue: "3"; pages: "385-390"
COMMENT     Sequence Label: UAS-Cre-T2A-miRFP670
FEATURES             Location/Qualifiers
     source          1..5263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             188..217
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             233..286
                     /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             287..1234
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     complement(1268..1285)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1274..1481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1505..1626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1516..1535)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1570..1589
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1874..1891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2045..2633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2125..2144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2833..3624,3625..3693))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3798..3846
                     /note="synthetic polyadenylation signal"
     misc_feature    3860..3951
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3900..3919
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    4036..4130
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        4149..4387
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     4334..4355
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      4407..4416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4413..5263,1..178)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
ORIGIN
        1 ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga gatatggccc gcgctggagt
       61 ttcaataccg gagatcatgc aagctggtgg ctggaccaat gtaaatattg tcatgaacta
      121 tatccgtaac ctggatagtg aaacaggggc aatggtgcgc ctgctggaag atggcgatgg
      181 accggtggaa caaaaactta tttctgaaga agatctgggg cccggcagtg gagagggcag
      241 aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg tagcaggtca
      301 tgcctctggc agccccgcat tcgggaccgc ctctcattcg aattgcgaac atgaagagat
      361 ccacctcgcc ggctcgatcc agccgcatgg cgcgcttctg gtcgtcagcg aacatgatca
      421 tcgcgtcatc caggccagcg ccaacgccgc ggaatttctg aatctcggaa gcgtactcgg
      481 cgttccgctc gccgagatcg acggcgatct gttgatcaag atcctgccgc atctcgatcc
      541 caccgccgaa ggcatgccgg tcgcggtgcg ctgccggatc ggcaatccct ctacggagta
      601 ctgcggtctg atgcatcggc ctccggaagg cgggctgatc atcgaactcg aacgtgccgg
      661 cccgtcgatc gatctgtcag gcacgctggc gccggcgctg gagcggatcc gcacggcggg
      721 ttcactgcgc gcgctgtgcg atgacaccgt gctgctgttt cagcagtgca ccggctacga
      781 ccgggtgatg gtgtatcgtt tcgatgagca aggccacggc ctggtattct ccgagtgcca
      841 tgtgcctggg ctcgaatcct atttcggcaa ccgctatccg tcgtcgactg tcccgcagat
      901 ggcgcggcag ctgtacgtgc ggcagcgcgt ccgcgtgctg gtcgacgtca cctatcagcc
      961 ggtgccgctg gagccgcggc tgtcgccgct gaccgggcgc gatctcgaca tgtcgggctg
     1021 cttcctgcgc tcgatgtcgc cgtgccatct gcagttcctg aaggacatgg gcgtgcgcgc
     1081 caccctggcg gtgtcgctgg tggtcggcgg caagctgtgg ggcctggttg tctgtcacca
     1141 ttatctgccg cgcttcatcc gtttcgagct gcgggcgatc tgcaaacggc tcgccgaaag
     1201 gatcgcgacg cggatcaccg cgcttgagag ctaagctagc gaattctaac tagagctcgc
     1261 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1321 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1381 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1441 aagggggagg attgggaaga gaatagcagg catgctgggg aggtaccgct tcgagcagac
     1501 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1561 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1621 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1681 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     1741 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     1801 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     1861 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1921 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1981 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2041 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2101 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2161 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2221 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2281 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2341 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2401 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2461 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2521 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2581 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2641 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2701 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2761 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa
     2821 ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga tctgcctatt
     2881 tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc gtgagggctt
     2941 accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg cccccgattt
     3001 gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg ctactttgtc
     3061 cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag
     3121 tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg
     3181 tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat cacccatatt
     3241 atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta agttggccgc
     3301 ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca tgccatccgt
     3361 aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg
     3421 gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac atagcagtac
     3481 tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc
     3541 gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt cagcatcttt
     3601 tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg
     3661 aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat attattgaag
     3721 catttatcag ggttactagt acgtctctca aggataagta agtaatatta aggtacggga
     3781 ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     3841 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     3901 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tggcctaact
     3961 ggccggtacc tgagctcgct agcctcgagg atatcaagat ctggcctcgg cggccaagct
     4021 tgcatgcctg caggtcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta
     4081 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagactct
     4141 agcgagcgcc ggagtataaa tagaggcgct tcgtctacgg agcgacaatt caattcaaac
     4201 aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat aaacaagcgc agctgaacaa
     4261 gctaaacaat ctgcagtaaa gtgcaagtta aagtgaatca attaaaagta accagcaacc
     4321 aagtaaatca actgcaacta ctgaaatctg ccaagaagta attattgaat acaagaagag
     4381 aactctgaat agggaattgg gaattcgcca ccatggccaa tttactgacc gtacaccaaa
     4441 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
     4501 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
     4561 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
     4621 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc
     4681 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
     4741 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
     4801 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
     4861 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
     4921 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
     4981 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
     5041 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
     5101 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     5161 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     5221 gattgattta cggcgctaag gatgactctg gtcagagata cct
//
