LOCUS       sequence-328536-        3527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic Puromycin resistance cassette in Alpha1, lacks physical marker
            or attB site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3527)
  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 3527)
  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: pHSP70B-CP6-pac
FEATURES             Location/Qualifiers
     source          1..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        176..278
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             279..1094
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             279..1094
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(352..371)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1208..1663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1295..1314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1505..1526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1684..2135
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2083..2104
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     rep_origin      complement(join(3000..3527,1..61))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3080..3099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
       61 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgtgatgatc
      181 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
      241 ttgagttatc gagattttca ggagctaagg aagctaaaat gagccatatt caacgggaaa
      301 cgtcttgctc aaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      361 gggctcgcga taatgtcggg caatcaggtg cgacaatcta ccgattgtat gggaagcccg
      421 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
      481 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
      541 tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc
      601 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
      661 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
      721 gactcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
      781 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
      841 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
      901 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
      961 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1021 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1081 atgagttttt ctgatttttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac
     1141 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgatgataag ctgtcaaaca
     1201 tgatgttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1441 agtgggccag cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1621 aaatttaacg cgaattttaa caaaatatta acgcttacaa ttttaacgaa ttcgtctcag
     1681 gagctagaat cccaaaacaa actggttatt gtggtaggtc atttgtttgg cagaaagaaa
     1741 actcgagaaa tttctctggc cgttattcgt tattctctct tttctttttg ggtcgctccc
     1801 tctctgcact aatgctctct cactctgtca cacagtaaac ggcatactgc tctcgttggt
     1861 tcgagagagc gcgcctcgaa tgttcgcgaa aagagcgccg gagtataaat agaggcgctt
     1921 cgtctacgga gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc
     1981 taagcaaata aacaagcgca gctgaacaag ctaaacaatc tgcagtaaag tgcaagttaa
     2041 agtgaatcaa ttaaaagtaa ccagcaacca agtaaatcaa ctgcaactac tgaaatctgc
     2101 caagaagtaa ttattgaata caagaagaga actctcaata ctcgaatcca tgtgggagtt
     2161 tattcttgac acagatattt atgatataat aactgagtaa gcttaacata aggaggaaaa
     2221 caaaatgacc gagtataagc ccacggtgcg tctggccacc cgtgatgatg tgccccgtgc
     2281 cgtgcgaacc ctggctgctg ccttcgccga ttacccagcc acccgtcaca ccgtggaccc
     2341 agaccgtcac attgagcgtg tgaccgagct ccaggagctg ttcctgacac gcgtgggcct
     2401 ggatatcggc aaggtctggg tggccgatga tggagctgcc gtggccgtgt ggaccacccc
     2461 cgagagcgtg gaggctggag ccgtgttcgc cgagattgga ccccgtatgg ccgagctgag
     2521 tggctcccgt ctggctgccc agcagcagat ggagggcctg ctggccccac accgtccaaa
     2581 ggagcccgtc tggttcctgg ccaccgtggg cgtgagcccc gaccaccagg gcaagggcct
     2641 gggcagtgcc gtggtgctgc caggagtgga agctgccgag cgtgctggag tgccagcctt
     2701 cctggaaacc agtgccccac gcaatctgcc cttctacgag cgtctgggct tcaccgtgac
     2761 ggccgatgtg gaggtgcccg agggcccacg cacctggtgt atgacccgta agccaggagc
     2821 ctaagcttgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttccgct
     2881 gtcatgagac gaattctgac acggttatcc acagaatcag gggataacgc aggaaagaac
     2941 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3001 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3061 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3121 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3181 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3241 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3301 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3361 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3421 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3481 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgc
//
