LOCUS       sequence-323278-        3499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1-147 AA of GAL4 DNA binding domain, requires assembly with TL2-TL4 
            linker and TL5 activation domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3499)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3499)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pGal4BD 1-147
FEATURES             Location/Qualifiers
     source          1..3499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     408..427
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        408..426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      457..466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      461..470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             505..945
                     /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"
     promoter        complement(1048..1066)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1049..1066)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1084..1100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1084..1100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1097..1119)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1108..1124
                     /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(1132..1138,1139..1156,1157..1162))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1177..1198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1315..1332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1486..2074)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1566..1585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2245..3036,3037..3105))
                     /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     2868..2887
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3106..3210)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3288..3499,1..244))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3425..3446)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gctggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
       61 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      121 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      181 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      241 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
      421 tatagggcga attgggcccg acgtcgcatg ctcccggccg ccatggcggc cgcgggaatt
      481 cgattgggcg atgagtggtc tcgaatgaag ctgctgagta gtattgaaca agcgtgtgac
      541 atatgccgac ttaagaagct gaagtgcagc aaggaaaagc cgaagtgcgc caaatgtctt
      601 aagaacaatt gggagtgccg gtactcgcca aagacgaagc gaagcccttt gacacgcgct
      661 catttgactg aagtggagag tcgcctggag cgccttgagc agttgttcct cctcattttt
      721 ccccgtgaag acctcgacat gatactgaag atggatagtc tccaagatat taaggccctg
      781 ctcactggcc tgtttgttca ggataacgtc aacaaggatg ccgtgacgga ccgccttgcc
      841 tcggtggaga cagatatgcc acttaccttg cggcagcatc gcatctcggc taccagcagt
      901 agtgaagaaa gttccaataa gggtcaacgg caactgacgg tatcgagtgc gagaccactc
      961 atcgcccaat cactagtgaa ttcgcggccg cctgcaggtc gaccatatgg gagagctccc
     1021 aacgcgttgg atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
     1081 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1141 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1201 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1261 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1321 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1381 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1441 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1501 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1561 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1621 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1681 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1741 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1801 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1861 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1921 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1981 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2041 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2101 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2161 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2221 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2281 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2341 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgcgacc cacgctcacc
     2401 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2461 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2521 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2581 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2641 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2701 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2761 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2821 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2881 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2941 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3001 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3061 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3121 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3181 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgatgc
     3241 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     3301 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3361 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3421 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3481 aaaaaccgtc tatcagggc
//
