LOCUS       sequence-328538-        3953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic Hygromycin 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 3953)
  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 3953)
  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-hph
FEATURES             Location/Qualifiers
     source          1..3953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     236..257
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             378..1403
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     rep_origin      complement(1579..2167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1659..1678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2282..2384
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2385..3200
                     /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             2385..3200
                     /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(2458..2477)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3314..3769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3401..3420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3611..3632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        join(3790..3953,1..288)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
ORIGIN
        1 tgctctcgtt ggttcgagag agcgcgcctc gaatgttcgc gaaaagagcg ccggagtata
       61 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
      121 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta
      181 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac
      241 tactgaaatc tgccaagaag taattattga atacaagaag agaactctca atactcgaat
      301 ccatgtggga gtttattctt gacacagata tttatgatat aataactgag taagcttaac
      361 ataaggagga aaacaaaatg aagaagcccg agctgacggc caccagcgtt gagaagttct
      421 tgattgagaa gtttgatagc gtgtccgacc tgatgcagct gagcgagggc gaggaaagcc
      481 gtgccttcag ctttgatgtg ggaggccgtg gatatgtgct gcgtgtgaac agctgtgccg
      541 atggcttcta caaggaccgt tatgtgtatc gccatttcgc cagtgctgcc ctgccaatcc
      601 ccgaggtgct ggatatcggc gagttcagcg agagcctgac ctactgcatt agccgtcgtg
      661 cccagggcgt gaccctgcaa gacctgcccg aaaccgagct gccagccgtg ctccagccag
      721 tggctgaggc tatggatgcc attgctgctg ccgacctgag ccagaccagt ggcttcggcc
      781 cattcggacc acaaggcatc ggccagtata ccacctggcg tgatttcatc tgtgccattg
      841 ccgacccaca cgtctaccac tggcagaccg tgatggatga taccgtgtcg gccagcgtgg
      901 cccaggccct ggacgagctg atgctgtggg ccgaggattg cccagaggtc cgtcacctgg
      961 tgcacgccga tttcggcagc aacaatgtgc tgaccgataa cggccgtatc acggccgtga
     1021 ttgattggag cgaggctatg ttcggcgata gccagtatga ggtggccaat atcttcttct
     1081 ggcgtccctg gctggcctgt atggaacagc agacccgtta ctttgagcgt cgccatccag
     1141 agctggctgg aagcccacgc ctgcgtgcct atatgctgcg tatcggcctg gaccagctct
     1201 accagagcct ggtggatggc aactttgatg atgctgcctg ggctcaggga cgctgcgacg
     1261 ccattgtgcg tagtggagct ggaaccgtgg gacgcaccca gattgcccgt cgctcggctg
     1321 ccgtgtggac cgatggatgc gtggaggtgc tggccgatag tggaaatcgc cgtccaagca
     1381 cccgtccccg tgccaaggag taagcttggc aataaaaaga cagaataaaa cgcacgggtg
     1441 ttgggtcgtt tgttccgctg tcatgagacg aattctgaca cggttatcca cagaatcagg
     1501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgtg
     2281 atgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     2341 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatgagc catattcaac
     2401 gggaaacgtc ttgctcaagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2461 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctaccga ttgtatggga
     2521 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2581 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2641 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatccca gggaaaacag
     2701 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2761 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2821 tatttcgact cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2881 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2941 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3001 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3061 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     3121 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     3181 tgctcgatga gtttttctga ttttttttaa ggcagttatt ggtgccctta aacgcctggt
     3241 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgat gataagctgt
     3301 caaacatgat gttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3541 tcacgtagtg ggccagcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3661 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3721 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttt aacgaattcg
     3781 tctcaggagc tagaatccca aaacaaactg gttattgtgg taggtcattt gtttggcaga
     3841 aagaaaactc gagaaatttc tctggccgtt attcgttatt ctctcttttc tttttgggtc
     3901 gctccctctc tgcactaatg ctctctcact ctgtcacaca gtaaacggca tac
//
