LOCUS       sequence-328541-        4050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic 5-fluorocytosine sensitivity cassette in Alpha2, lacks 
            physical marker or attB site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4050)
  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 4050)
  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-FCU1
FEATURES             Location/Qualifiers
     source          1..4050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(103..122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     313..334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        497..948
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     896..917
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     rep_origin      complement(2331..2919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2411..2430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3034..3136
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3137..3952
                     /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             3137..3952
                     /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(3210..3229)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
       61 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      121 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      181 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      241 gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      301 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      361 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      421 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt ttaacaagct
      481 tcgtctcagt caggagctag aatcccaaaa caaactggtt attgtggtag gtcatttgtt
      541 tggcagaaag aaaactcgag aaatttctct ggccgttatt cgttattctc tcttttcttt
      601 ttgggtcgct ccctctctgc actaatgctc tctcactctg tcacacagta aacggcatac
      661 tgctctcgtt ggttcgagag agcgcgcctc gaatgttcgc gaaaagagcg ccggagtata
      721 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
      781 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atctgcagta
      841 aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat caactgcaac
      901 tactgaaatc tgccaagaag taattattga atacaagaag agaactctca atactcgaat
      961 ccatgtggga gtttattctt gacacagata tttatgatat aataactgag taagcttaac
     1021 ataaggagga aaacaaaatg gtgaccggcg gcatggccag caagtgggat caaaagggca
     1081 tggatattgc ctatgaggag gccgccttgg gctacaagga gggcggcgtc cccattggcg
     1141 gctgcttgat caacaacaag gatggcagcg tgctgggacg cggccacaac atgcgcttcc
     1201 agaagggcag cgccaccttg cacggcgaga tcagcaccct ggagaactgc ggacgcttgg
     1261 agggcaaggt gtacaaggat accaccctgt acaccaccct gagcccctgc gatatgtgca
     1321 ccggcgccat catcatgtac ggcatccccc gctgcgtggt gggcgagaac gtgaacttca
     1381 agagcaaggg cgagaagtat ctgcaaaccc gcggacacga ggtggtggtg gtggatgatg
     1441 agcgatgcaa aaagatcatg aagcagttca tcgatgagcg cccccaggat tggttcgagg
     1501 atattggcga ggccagcgag cccttcaaga acgtgtatct gctgccccag accaaccaac
     1561 tgctgggcct gtacaccatc atccgcaaca agaacaccac ccgcccagat ttcatcttct
     1621 attgtgaccg catcatccgc ctgctggtcg aggagggcct gaaccacctg cccgtgcaga
     1681 agcagatcgt cgaaaccgat accaacgaga acttcgaggg cgttagcttc atgggcaaga
     1741 tctgcggcgt gagcatcgtg cgtgccggcg agagcatgga gcagggcctg cgcgattgct
     1801 gccgcagcgt gcgcatcggc aagatcctga tccagcgcga cgaggaaacc gccctgccca
     1861 agctgtttta cgagaagctg cccgaggata tttccgagcg ctacgtgttc ctgctcgatc
     1921 ccatgctggc caccggcggc agcgccatca tggccaccga ggtgctgatc aagcgcggcg
     1981 tgaagcccga gcgcatctac tttctgaacc tgatttgcag caaggagggc attgagaagt
     2041 accacgccgc cttcccggag gtccgcatcg tgaccggagc cttggatcgc ggattggatg
     2101 agaacaaata tctggtgccc ggcctgggcg atttcggcga tcgctactac tgcgtgtaag
     2161 cttggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt ccgcttgaga
     2221 cgaagcttga cacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2281 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2341 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2401 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2461 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2521 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2581 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2641 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2701 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2761 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2821 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2881 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2941 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3001 tcaaaaagga tcttcaccta gatccttttg tgatgatcgg cacgtaagag gttccaactt
     3061 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     3121 agctaaggaa gctaaaatga gccatattca acgggaaacg tcttgctcaa ggccgcgatt
     3181 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3241 atcaggtgcg acaatctacc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     3301 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3361 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3421 gttactcacc actgcgatcc cagggaaaac agcattccag gtattagaag aatatcctga
     3481 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3541 tgtttgtaat tgtcctttta acagcgatcg cgtatttcga ctcgctcagg cgcaatcacg
     3601 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3661 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtagtcac
     3721 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3781 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3841 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3901 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct gatttttttt
     3961 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
     4021 ggatgaatgg cagaaattcg atgataagct
//
