LOCUS       sequence-323395-        3311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic Blasticidin 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 3311)
  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 3311)
  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-bsr
FEATURES             Location/Qualifiers
     source          1..3311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..472
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     420..441
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     rep_origin      complement(1160..1748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1240..1259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1863..1965
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1966..2781
                     /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             1966..2781
                     /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(2039..2058)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2921..3301
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3008..3027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3218..3239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacgaattc gtctcaggag ctagaatccc aaaacaaact ggttattgtg gtaggtcatt
       61 tgtttggcag aaagaaaact cgagaaattt ctctggccgt tattcgttat tctctctttt
      121 ctttttgggt cgctccctct ctgcactaat gctctctcac tctgtcacac agtaaacggc
      181 atactgctct cgttggttcg agagagcgcg cctcgaatgt tcgcgaaaag agcgccggag
      241 tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca
      301 cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatctgc
      361 agtaaagtgc aagttaaagt gaatcaatta aaagtaacca gcaaccaagt aaatcaactg
      421 caactactga aatctgccaa gaagtaatta ttgaatacaa gaagagaact ctcaatactc
      481 gaatccatgt gggagtttat tcttgacaca gatatttatg atataataac tgagtaagct
      541 taacataagg aggaaaacaa aatgaaaacc ttcaacatca gccagcagga cctggaactg
      601 gtggaggtgg ctaccgagaa gattacgatg ctctacgagg ataacaagca ccacgtggga
      661 gctgccattc gcaccaagac gggcgagatt atcagtgccg tgcacattga ggcctacatc
      721 ggacgcgtga ccgtgtgtgc cgaggccatt gccattggca gtgccgtgag caacggccag
      781 aaggattttg ataccattgt ggccgtgcgt cacccatact ccgatgaggt ggaccgtagc
      841 atccgtgtcg tgtccccgtg tggaatgtgc cgtgagctga ttagcgatta cgcccccgat
      901 tgcttcgtgc tgattgagat gaacggcaag ctggtcaaga ccaccattga ggaactgatt
      961 cccctgaagt atacccgtaa ctaagcttgg caataaaaag acagaataaa acgcacgggt
     1021 gttgggtcgt ttgttccgct gtcatgagac gaattctgac acggttatcc acagaatcag
     1081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1561 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgt
     1861 gatgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1921 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag ccatattcaa
     1981 cgggaaacgt cttgctcaag gccgcgatta aattccaaca tggatgctga tttatatggg
     2041 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctaccg attgtatggg
     2101 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2161 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     2221 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca
     2281 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2341 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2401 gtatttcgac tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2461 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2521 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2581 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2641 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2701 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2761 atgctcgatg agtttttctg atttttttta aggcagttat tggtgccctt aaacgcctgg
     2821 ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga tgataagctg
     2881 tcaaacatga tgttcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3301 gcctattggt t
//
