LOCUS       sequence-323284-        4366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4 inhibitor coding sequence (CDS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4366)
  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 4366)
  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: pGAL80
FEATURES             Location/Qualifiers
     source          1..4366
                     /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"
     promoter        complement(1915..1933)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1916..1933)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1951..1967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1951..1967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1964..1986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1975..1991
                     /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(1999..2005,2006..2023,2024..2029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2044..2065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2182..2199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2353..2941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2433..2452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3112..3903,3904..3972))
                     /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     3735..3754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3973..4077)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4155..4366,1..244))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4292..4313)
                     /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 tcgaatggat tacaacaaaa ggagtagtgt gagtacggtg
      541 ccgaatgctg ctcccattcg cgtgggcttc gtgggattga acgcggctaa gggttgggcc
      601 attaaaacgc attatccagc catactgcag ctgagctccc agttccaaat aacagcgctt
      661 tattccccca agatcgagac atccattgcg acaattcaga ggctgaagtt gtccaacgcc
      721 acagctttcc caacactcga gagcttcgcc tcgagctcca cgatcgatat gattgtgatc
      781 gcaatccagg tggcttccca ctacgaggtg gtaatgcccc tgctcgagtt ttcgaagaac
      841 aatcctaatc tgaagtattt gtttgtggaa tgggcactgg cctgctcgct cgatcaggcg
      901 gagtcgatct ataaggccgc tgcagagcgg ggcgtgcaaa caataatcag tctgcaggga
      961 cgaaagagcc cgtacatttt gcgggcaaaa gagcttatct cccagggcta catcggcgat
     1021 ataaatagca tcgagattgc aggtaacgga ggttggtacg gttacgagcg gccggtgaaa
     1081 agcccgaaat acatttacga gatcggaaac ggagttgatc tggtgaccac cacgttcggt
     1141 cacacgatag atatattgca gtacatgacc agttcgtact tcagccgtat caatgccatg
     1201 gtgtttaaca acattccaga acaggagctc attgatgaga ggggcaatcg cctgggccag
     1261 cgggtcccaa agactgtccc agatcatctc ttgttccagg gcacgctgct caacggaaat
     1321 gttcctgtgt cctgttcctt taagggaggc aagcctacga aaaagttcac caagaatctg
     1381 gtcatagata tccatggtac aaagggtgat ctgaagctgg agggtgacgc tggatttgcg
     1441 gaaatctcga atctcgttct gtattattcg ggcacccgcg ccaatgattt tcccctggcc
     1501 aatggtcagc aggcaccctt ggaccccggc tacgacgcag gcaaggagat tatggaggtg
     1561 tatcacctcc gaaactacaa cgcgatcgtt ggtaatatac accggctcta ccagagtatc
     1621 agcgatttcc acttcaacac caagaaaatc cccgagcttc catcgcagtt cgtaatgcaa
     1681 ggcttcgatt tcgaaggctt tcccaccctg atggatgcct tgatcttgca tcgcctgatt
     1741 gaaagtgtct acaagtcgaa tatgatgggc agtaccctga atgtttccaa catctcccac
     1801 tactcgcttt aagcttcgag accactcatc gcccaatcac tagtgaattc gcggccgcct
     1861 gcaggtcgac catatgggag agctcccaac gcgttggatg catagcttga gtattctata
     1921 gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1981 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2041 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2101 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2761 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3241 aatgataccg cgcgacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4081 atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4141 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4201 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4261 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4321 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggc
//
