LOCUS       sequence-328540-        3302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic Phleomycin 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 3302)
  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 3302)
  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-ble
FEATURES             Location/Qualifiers
     source          1..3302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..452
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     400..421
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     rep_origin      complement(1092..1680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1172..1191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1795..1897
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1898..2713
                     /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             1898..2713
                     /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(1971..1990)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2827..3282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2914..2933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3124..3145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctagaatccc aaaacaaact ggttattgtg gtaggtcatt tgtttggcag aaagaaaact
       61 cgagaaattt ctctggccgt tattcgttat tctctctttt ctttttgggt cgctccctct
      121 ctgcactaat gctctctcac tctgtcacac agtaaacggc atactgctct cgttggttcg
      181 agagagcgcg cctcgaatgt tcgcgaaaag agcgccggag tataaataga ggcgcttcgt
      241 ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa
      301 gcaaataaac aagcgcagct gaacaagcta aacaatctgc agtaaagtgc aagttaaagt
      361 gaatcaatta aaagtaacca gcaaccaagt aaatcaactg caactactga aatctgccaa
      421 gaagtaatta ttgaatacaa gaagagaact ctcaatactc gaatccatgt gggagtttat
      481 tcttgacaca gatatttatg atataataac tgagtaagct taacataagg aggaaaacaa
      541 aatggccaag ctgaccagtg ccgtgccagt gctgacggct cgtgatgtgg caggagccgt
      601 ggagttctgg accgaccgtc tgggcttcag tcgcgatttc gtggaggatg atttcgcagg
      661 cgtggtccgt gacgatgtga ccctgttcat ctcggccgtg caggaccagg tggtgcccga
      721 taacaccctg gcctgggtct gggtccgtgg cctggatgag ctctacgccg agtggagcga
      781 ggtggtgtcc accaacttcc gagatgccag tggaccagca atgaccgaga ttggcgagca
      841 gccgtgggga cgcgagttcg ccctccgtga cccagcaggc aactgcgtgc acttcgtggc
      901 cgaggagcag gactaagctt ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
      961 gtttgttccg ctgtcatgag acgaattctg acacggttat ccacagaatc aggggataac
     1021 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gtgatgatcg
     1801 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     1861 tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac
     1921 gtcttgctca aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     1981 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctac cgattgtatg ggaagcccga
     2041 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2101 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2161 ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca
     2221 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2281 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2341 actcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2401 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2461 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2521 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2581 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2641 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2701 tgagtttttc tgattttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     2761 cctgaataag tgataataag cggatgaatg gcagaaattc gatgataagc tgtcaaacat
     2821 gatgttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2881 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2941 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3001 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3061 gtgggccagc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3241 aatttaacgc gaattttaac aaaatattaa cgcttacaat tttaacgaat tcgtctcagg
     3301 ag
//
