LOCUS       sequence-357481-        6974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for conversion of KO attP Drosophila lines into 
            ShineGal4 drivers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6974)
  AUTHORS   di Pietro F, Herszterg S, Huang A, Bosveld F, Alexandre C, Sancere 
            L, Pelletier S, Joudat A, Kapoor V, Vincent JP, Bellaiche Y
  TITLE     Rapid and robust optogenetic control of gene expression in 
            Drosophila.
  JOURNAL   Dev Cell. 2021 Dec 20;56(24):3393-3404.e7. doi: 
            10.1016/j.devcel.2021.11.016. Epub 2021 Dec 7.
  PUBMED    34879263
REFERENCE   2  (bases 1 to 6974)
  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"; doi: 
            "10.1016/j.devcel.2021.11.016"; journalName: "Dev Cell"; date: 
            "2021-12-20- 20"; volume: "56"; issue: "24"; pages: "3393-3404.e7"
COMMENT     Sequence Label: pRIV-Gal4DBD:2xnMagHigh1
FEATURES             Location/Qualifiers
     source          1..6974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(460..490,491..692)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    726..759
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             800..1240
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    2965..2998
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3181..3891
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3326..3344)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3594..3613
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4011..4132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4048..4067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4102..4121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4149..4703,4704..4734)
                     /label=P element 5' end
     primer_bind     complement(4753..4769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4753..4769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4766..4788)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4777..4793
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4801..4807,4808..4825,4826..4831))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4846..4867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4984..5001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5155..5743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5235..5254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5914..6705,6706..6774))
                     /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     6537..6556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6775..6879)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6947..6965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgtcga ccgaagtctt agagccagat
      421 atgcgagcac ccggaagctc acgatgagaa tggccagacc atgatgaaat aacataaggt
      481 ggtcccgtcg gcaagagaca tccacttaac gtatgcttgc aataagtgcg agtgaaagga
      541 atagtattct gagtgtcgta ttgagtctga gtgagacagc gatatgattg ttgattaacc
      601 cttagcatgt ccgtggggtt tgaattaact cataatatta attagacgaa attattttta
      661 aagttttatt tttaataatt tgcgagtacg caaagctatg catgtaatga attgccgcgg
      721 tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actcccgtac gacgaattct
      781 ccggagcggc cgctctagaa tgaaactgtt gagttccata gagcaggcat gcgatatctg
      841 ccgtttgaaa aaactgaaat gttcgaagga gaaaccaaag tgtgcgaaat gtctcaagaa
      901 caactgggaa tgccgatact cgccgaaaac gaaacgatcg ccgctgacgc gggctcatct
      961 cacagaggtg gaatcgcgtc tcgaacgatt ggagcaactc tttctgttga tctttccgag
     1021 ggaagacctg gacatgattc tgaagatgga tagcttgcag gatataaagg cgctcctcac
     1081 cggcctcttt gtgcaggata atgtaaacaa agatgctgta acagatcgcc tggcctccgt
     1141 ggagactgac atgccgttga cgttgaggca gcatcgcata tccgcgactt cgagttcgga
     1201 ggaatccagt aacaaaggtc aacgacagct cactgtttcg ggaggcggag gctccggtgg
     1261 tggtggaagt ggcggtggcg gatcccatac cctgtacgcg cccggaggtt atgacattat
     1321 gggttatctc gatcagatcg gaaaccgtcc caatccgcag gttgagctgg gaccagtgga
     1381 tacgtcctgt gctttgattc tctgtgatct gaaacagaaa gataccccta ttgtttatgc
     1441 aagtgaggcc tttctgtata tgaccggtta ctccaacgcg gaggtgttgg gccggaattg
     1501 tcgttttctg cagtcgccag acggaatggt caaacctaaa tccacacgca agtatgtgga
     1561 cagcaatact ataaatacga ttcgaaaggc aatagaccgc aacgctgaag tccaggtgga
     1621 ggtcgttaat ttcaagaaga atggacaaag gttcgtcaac ttcttgacga tcatcccggt
     1681 ccgtgacgag acaggtgagt atcgatatag tatgggcttt caatgcgaaa cggaaggtgg
     1741 aggtggcagt ggaggaggag gaagcggtgg cggtggcagt cataccttgt atgcaccagg
     1801 cggttacgac ataatgggct atttggacca aatcggaaac aggccgaacc cccaagtcga
     1861 gctcggacca gtagatacaa gctgcgcgct catcttgtgt gatttgaaac aaaaggatac
     1921 ccctatagtg tatgccagcg aagcattcct ctacatgaca ggctacagca acgcagaagt
     1981 tctgggaagg aactgtcgat ttctccagtc gccagacggc atggtcaaac cgaaatcgac
     2041 tcggaagtat gtagatagta atacaatcaa cacgatacgt aaagccattg acaggaatgc
     2101 ggaagtacaa gttgaggttg ttaattttaa gaaaaatggc cagaggtttg ttaactttct
     2161 cacaataata cctgtgaggg acgaaaccgg cgagtacagg tacagcatgg gattccaatg
     2221 tgagacggaa taatctagaa ggcctggcgc gccgctagca tgaatcgttt ttaaaataac
     2281 aaatcaattg ttttataata ttcgtacgat tctttgatta tgtaataaaa tgtgatcatt
     2341 aggaagatta cgaaaaatat aaaaaatatg agttctgtgt gtataacaaa tgctgtaaac
     2401 gccacaattg tgtttgttgc aaataaaccc atgattattt gattaaaatt gttgttttct
     2461 ttgttcatag acaatagtgt gttttgccta aacgtgtact gcataaactc catgcgagtg
     2521 tatagcgagc tagtggctaa cgcttgcccc accaaagtag attcgtcaaa atcctcaatt
     2581 tcatcaccct cctccaagtt taacatttgg ccgtcggaat taacttctaa agatgccaca
     2641 taatctaata aatgaaatag agattcaaac gtggcgtcat cgtccgtttc gaccatttcc
     2701 gaaaagaact cgggcataaa ctctatgatt tctctggacg tggtgttgtc gaaactctca
     2761 aagtacgcag tcaggaacgt gcgcgacatg tcgtcgggaa actcgcgcgg aaacatgttg
     2821 ttgtaaccga acgggtccca tagcgccaaa accaaatctg ccagcgtcaa tagaatggca
     2881 cgatgccgac aatggagctg gcttggatag cgattcgagt taacggatcc actagtgctc
     2941 gagtggcgcg ccggtaccac cggtataact tcgtataatg tatgctatac gaagttatgc
     3001 ccggatctaa ttcaattaga gactaattca attagagcta attcaattag cgccggagta
     3061 taaatagagg cgcttcgtct acggagcgac aattcaattc aaacaagcaa agtgaacacg
     3121 tcgctaagcg aaagctaagc aaataaacaa gcgcagctga acaagctaaa caatcggccc
     3181 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     3241 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     3301 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     3361 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     3421 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     3481 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     3541 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     3601 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     3661 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     3721 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     3781 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     3841 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agcgccgcga
     3901 ctctgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3961 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4021 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4081 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tagctggatc
     4141 ctaattcggc tgctgctcta aacgacgcat ttcgtactcc aaagtacgaa ttttttccct
     4201 caagctctta ttttcattaa acaatgaaca ggacctaacg cacagtcacg ttattgttta
     4261 cataaatgat tttttttact attcaaactt actctgtttg tgtactccca ctggtatagc
     4321 cttcttttat cttttctggt tcaggctcta tcactttact aggtacggca tctgcgttga
     4381 gtcgcctcct tttaaatgtc tgaccttttg caggtgcagc cttccactgc gaatcattaa
     4441 agtgggtatc acaaatttgg gagttttcac caaggctgca cccaaggctc tgctcccaca
     4501 attttctctt aatagcacac ttcggcacgt gaattaattt tactccagtc acagctttgc
     4561 agcaaaattt gcaatatttc attttttttt attccacgta agggttaatg ttttcaaaaa
     4621 aaaattcgtc cgcacacaac ctttcctctc aacaagcaaa cgtgcactga atttaagtgt
     4681 atacttcggt aagcttcggc tttcgacggg accaccttat gttatttcat catgacgcgt
     4741 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4801 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4861 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4921 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4981 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5041 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5881 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5941 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6001 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6061 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6121 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6181 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6241 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6301 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6361 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6421 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6481 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6541 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6601 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6661 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6721 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6781 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6841 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6901 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6961 gaggcccttt cgtc
//
