LOCUS       sequence-328891-        3722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic G418 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 3722)
  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 3722)
  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-nptII
FEATURES             Location/Qualifiers
     source          1..3722
                     /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(1512..2100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1592..1611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2215..2317
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2318..3133
                     /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             2318..3133
                     /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(2391..2410)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3247..3702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3334..3353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3544..3565
                     /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 aatgattgag caggatggac tgcacgctgg ctcgccagct gcctgggtgg agcgtctgtt
      601 cggctacgat tgggcccagc agaccatcgg ctgctccgat gctgccgtgt tccgtctgtc
      661 ggcccaggga cgccccgtgc tgttcgtcaa gaccgacctg agtggagccc tgaacgagct
      721 ccaggatgag gcagctcgtc tgagctggct ggccacaaca ggagtgccgt gtgctgccgt
      781 gctggatgtg gtgaccgaag ctggacgcga ttggctgctg ctgggcgagg tgccaggcca
      841 ggacctgctg tccagccatc tggccccagc cgagaaggtg tccattatgg ccgatgctat
      901 gcgtcgcctg cacaccctgg acccagccac ctgccccttt gaccaccagg ccaagcaccg
      961 tattgagcgt gcccgaaccc gtatggaggc tggactggtg gaccaggatg acctggatga
     1021 ggagcaccag ggcctggctc cagccgagct gtttgcccgt ctgaaggccc gtatgcccga
     1081 tggcgaggac ctggtggtga cccacggcga cgcctgcctg cccaacatta tggtggagaa
     1141 cggacgcttc agtggcttca ttgattgtgg acgcctgggc gtggccgacc gttaccagga
     1201 tatcgccctg gccacccgag atatcgccga ggagctggga ggcgagtggg ccgaccgatt
     1261 cctggtgctc tacggcattg cagccccaga tagccagcgt attgccttct accgtctgct
     1321 ggacgagttc ttctaagctt ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     1381 gtttgttccg ctgtcatgag acgaattctg acacggttat ccacagaatc aggggataac
     1441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1921 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gtgatgatcg
     2221 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     2281 tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac
     2341 gtcttgctca aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2401 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctac cgattgtatg ggaagcccga
     2461 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2521 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2581 ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca
     2641 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2701 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2761 actcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2821 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2881 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2941 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3001 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3061 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3121 tgagtttttc tgattttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     3181 cctgaataag tgataataag cggatgaatg gcagaaattc gatgataagc tgtcaaacat
     3241 gatgttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3301 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3361 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3421 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3481 gtgggccagc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3541 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3601 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3661 aatttaacgc gaattttaac aaaatattaa cgcttacaat tttaacgaat tcgtctcagg
     3721 ag
//
