LOCUS       sequence-431463-        3192 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 3192)
  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 3192)
  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: AA365
FEATURES             Location/Qualifiers
     source          1..3192
                     /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    461..494
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    507..601
                     /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    617..650
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    663..757
                     /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        781..1019
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     966..987
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     complement(1102..1118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1102..1118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1115..1137)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1126..1142
                     /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(1150..1156,1157..1174,1175..1180))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1195..1216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1333..1350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1504..2092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1584..1603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2270..3079)
                     /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     2993..3012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3071..3090
                     /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 cacactagta gtaccagctt ataacttcgt ataatgtatg
      481 ctatacgaag ttatctgcag gcaggtcgga gtactgtcct ccgagcggag tactgtcctc
      541 cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga
      601 gcggagactc ccatggataa cttcgtataa tgtatgctat acgaagttat ggatccgcag
      661 gtcggagtac tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc
      721 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agactcgtcg acagcgagcg
      781 ccggagtata aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag
      841 tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca
      901 atctgcagta aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat
      961 caactgcaac tactgaaatc tgccaagaag taattattga atacaagaag agaactctga
     1021 atagggaatt ggaggagcgt cttcatcgga tcccgggccc gtcgactgca gaggcctgca
     1081 tgcaagcttg gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1141 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1201 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1261 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1321 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1381 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1441 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1501 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1561 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1621 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1681 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1741 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1801 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1861 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1921 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1981 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2041 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2101 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2161 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2221 taaatcaagc ccaatctgaa taatgttaca accaattaac caattctgat tagaaaaact
     2281 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2341 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2401 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2461 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2521 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2581 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2641 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2701 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2761 cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2821 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2881 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2941 catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3001 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt
     3061 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     3121 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg
     3181 gccagagctg ca
//
