LOCUS       sequence-165529-        4596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor HR template carrying a 2,080 bp fragment of the Drosophila 
            melanogaster white[coffee] allele and silent mutations conferring 
            resistance to white sgRNAs-1, -2, -3, and -4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4596)
  AUTHORS   Ge DT, Tipping C, Brodsky MH, Zamore PD
  TITLE     Rapid Screening for CRISPR-Directed Editing of the Drosophila Genome
            Using white Co-conversion.
  JOURNAL   G3 (Bethesda). 2016 Aug 26. pii: g3.116.032557. doi: 
            10.1534/g3.116.032557.
  PUBMED    27543296
REFERENCE   2  (bases 1 to 4596)
  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: "G3 
            (Bethesda). 2016 Aug 26. pii: g3.116.032557. doi: 
            10.1534/g3.116.032557."
COMMENT     Sequence Label: pUC57-white[coffee]
FEATURES             Location/Qualifiers
     source          1..4596
                     /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"
     primer_bind     complement(2506..2522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2506..2522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2519..2541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2530..2546
                     /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(2554..2560,2561..2578,2579..2584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2599..2620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2737..2754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2908..3496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2988..3007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3674..4483)
                     /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     4397..4416
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4475..4494
                     /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 ccagtgaatt cgagctcttt gacaagatcc
      421 ttctgatggc cgagggcagg gtagctttct tgggcactcc cagcgaagcc gtcgacttct
      481 tttcctagtg agttcgatgt gtttattaag ggtatctagt attacataac atctcaactc
      541 ctatccagcg tgggtgccca gtgtcctacc aactacaatc cggcggactt ttacgtacag
      601 gtgttggccg ttgtgcccgg acgggagatc gagtcccgtg atcggatcgc caagatatgc
      661 gacaattttg ccattagcaa agtagcccgg gatatggagc agttgttggc caccaaaaat
      721 ctggagaagc cactggagca gccggagaat gggtacacct acaaggccac ctggttcatg
      781 cagttccggg cggtcctgtg gcgatcctgg ctgtcggtgc tcaaggaacc actcctcgta
      841 aaagtgcgac ttattcagac aacggtgagt ggttccagtg gaaacaaatg atataacgct
      901 tacaattctt ggaaacaaat tcgctagatt ttagatagaa ttgcctgatt ccacaccctt
      961 cttagttttt ttcaatgaga tgtatagttt atagttttgc agaagataaa taaatttcat
     1021 ttaactcgcg aatattaatg agatgcgagt aacattttaa tttgcagatg gttgccatct
     1081 tgattggcct catctttttg ggccaacaac tcacgcaagt gggtgtgatg aatatcaacg
     1141 gagccatctt cctcttcctg accaacatga cctttcaaaa cgtctttgcc acgataaatg
     1201 taagtcatgt ttagaataca tttgcatttc aataatttac taactttcta atgaatcgat
     1261 tcgatttagg tgttcacctc agagctgcca gtttttatga gggaggcccg aagtcgactt
     1321 tatcgctgtg acacatactt tctgggcaaa acgattgccg aattgccgct ttttctcaca
     1381 gtgccactgg tcttcacggc gattgcctat ccgatgatcg gactgcgggc cggagtgctg
     1441 cacttcttca actgcctggc gctggtcact ctggtggcca atgtgtcaac gtccttcgga
     1501 tatctaatat cctgcgccag ctcctcgacc tcgatggcgc tgtctgtggg tccgccggtt
     1561 atcataccat tcctgctctt tggcgaattc ttcttgaact cgggctcggt gccagtttac
     1621 ctcaaatggt tgtcgtacct ctcatggttc cgttacgcca acgagggtct gctgattaac
     1681 caatgggcgg acgtggagcc gggcgaaatt agctgcacat cgtcgaacac cacgtgcccc
     1741 agttcgggca aggtcatcct ggagacgctt aacttctcgg ccgccgatct gccgctggac
     1801 tacgtgggtc tggccattct catcgtgagc ttccgggtgc tcgcatatct ggctctaaga
     1861 cttcgggccc gacgcaagga gtagccgaca tatatccgaa ataactgctt gttttttttt
     1921 taccattatt accatcgtgt ttactgttta ttgccccctc aaaaagctaa tgtaattata
     1981 tttgtgccaa taaaaacaag atatgaccta tagaatacaa gtatttcccc ttcgaacatc
     2041 cccacaagta gactttggat ttgtcttcta accaaaagac ttacacacct gcatacctta
     2101 catcaaaaac tcgtttatcg ctacataaaa caccgggata tattttttat atacaaactt
     2161 ttcaaatcgc gcgccctctt cataattcac ctccaccaca ccacgtttcg tagttgctct
     2221 ttcgctgtct cccacccgct ctccgcaaca cattcacctt ttgttcgacg accttggagc
     2281 gactgtcgtt agttccgcgc gattcggttc gctcaaatgg ttccgagtgg ttcatttcgt
     2341 ctcaatagaa attagtaata aatatttgta tgtacaattt atttgctcca atatatttgt
     2401 atatatttcc ctcacagcta tatttattct aatttaatat tatgactttt taaggtaatt
     2461 ttttgtgacc tgttcggagt ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     2521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3601 ccttttaaat taaaaatgaa gttttaaatc aagcccaatc tgaataatgt tacaaccaat
     3661 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     3721 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     3781 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     3841 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     3901 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     3961 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     4021 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     4081 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     4141 ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga
     4201 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     4261 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     4321 catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac
     4381 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     4441 aatttaatcg cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac
     4501 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     4561 aacatcagag attttgagac acgggccaga gctgca
//
