LOCUS       sequence-323394-        3488 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 3488)
  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 3488)
  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..3488
                     /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(1337..1925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1417..1436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2040..2142
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2143..2958
                     /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             2143..2958
                     /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(2216..2235)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3098..3478
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3395..3416
                     /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 aatgaccgag tataagccca cggtgcgtct ggccacccgt
      601 gatgatgtgc cccgtgccgt gcgaaccctg gctgctgcct tcgccgatta cccagccacc
      661 cgtcacaccg tggacccaga ccgtcacatt gagcgtgtga ccgagctcca ggagctgttc
      721 ctgacacgcg tgggcctgga tatcggcaag gtctgggtgg ccgatgatgg agctgccgtg
      781 gccgtgtgga ccacccccga gagcgtggag gctggagccg tgttcgccga gattggaccc
      841 cgtatggccg agctgagtgg ctcccgtctg gctgcccagc agcagatgga gggcctgctg
      901 gccccacacc gtccaaagga gcccgtctgg ttcctggcca ccgtgggcgt gagccccgac
      961 caccagggca agggcctggg cagtgccgtg gtgctgccag gagtggaagc tgccgagcgt
     1021 gctggagtgc cagccttcct ggaaaccagt gccccacgca atctgccctt ctacgagcgt
     1081 ctgggcttca ccgtgacggc cgatgtggag gtgcccgagg gcccacgcac ctggtgtatg
     1141 acccgtaagc caggagccta agcttggcaa taaaaagaca gaataaaacg cacgggtgtt
     1201 gggtcgtttg ttccgctgtc atgagacgaa ttctgacacg gttatccaca gaatcagggg
     1261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1741 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgtgat
     2041 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     2101 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg
     2161 gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     2221 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctaccgatt gtatgggaag
     2281 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     2341 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     2401 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca
     2461 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2521 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     2581 tttcgactcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     2641 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2701 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2761 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2821 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2881 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2941 ctcgatgagt ttttctgatt ttttttaagg cagttattgg tgcccttaaa cgcctggttg
     3001 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgatga taagctgtca
     3061 aacatgatgt tcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3301 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3421 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3481 tattggtt
//
