LOCUS       sequence-431545-        3442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Pol 2 promoter) Gal4-induced expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3442)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 3442)
  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 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA366
FEATURES             Location/Qualifiers
     source          1..3442
                     /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"
     protein_bind    444..538
                     /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)"
     protein_bind    572..666
                     /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)"
     protein_bind    700..794
                     /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)"
     protein_bind    828..922
                     /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        947..1185
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     1132..1153
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     intron          1200..1266
                     /label=Mhc intron
                     /note="modified Drosophila myosin heavy chain intron 16 
                     (Pfeiffer et al., 2010)"
     primer_bind     complement(1352..1368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1352..1368)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1365..1387)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1376..1392
                     /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(1400..1406,1407..1424,1425..1430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1445..1466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1583..1600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1754..2342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1834..1853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2520..3329)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3243..3262
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3321..3340
                     /label=pENTR-R
                     /note="pENTR vectors, reverse 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 ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta caccggagta ctgtcctccg agcggagtac tgtcctccga
      481 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg
      541 gagactctag ccctagggca tgcctgcagg tcggagtact gtcctccgag cggagtactg
      601 tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc
      661 tccgagcgga gactctagcg ctagcgcatg cctgcaggtc ggagtactgt cctccgagcg
      721 gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag
      781 tactgtcctc cgagcggaga ctctagcact agtgcatgcc tgcaggtcgg agtactgtcc
      841 tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc
      901 gagcggagta ctgtcctccg agcggagact ctagcgacgt cgagcgccgg agtataaata
      961 gaggcgcttc gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta
     1021 agcgaaagct aagcaaataa acaagcgcag ctgaacaagc taaacaatct gcagtaaagt
     1081 gcaagttaaa gtgaatcaat taaaagtaac cagcaaccaa gtaaatcaac tgcaactact
     1141 gaaatctgcc aagaagtaat tattgaatac aagaagagaa ctctgaatag atctaaaagg
     1201 taggttcaac cactgatgcc taggcacacc gaaacgacta accctaattc ttatccttta
     1261 cttcaggcgg ccgcggctcg agaggagcgt cttcatcgga tcccgggccc gtcgactgca
     1321 gaggcctgca tgcaagcttg gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2461 aatgaagttt taaatcaagc ccaatctgaa taatgttaca accaattaac caattctgat
     2521 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     2581 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2641 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2701 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2761 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     2821 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     2881 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     2941 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     3001 tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
     3061 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     3121 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     3181 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
     3241 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     3301 ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3361 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     3421 tgagacacgg gccagagctg ca
//
