LOCUS       sequence-330498-        9315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xQUAS mCherry Mini-White CDS response vector.
            Contains an Alpha level GB2.0 entry point for addition of custom 
            assemblies utilizing blue-white bacterial colony screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9315)
  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 9315)
  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: pBlastR-5xQUAS-mCherry-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2966..2984
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     polyA_signal    3072..3206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3109..3128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3163..3182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4781..4802)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    5229..5298
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     5355..5374
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5568..5590
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5582..5599
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5583..5599
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5606..5625
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5606..5624
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5627..5683
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(5690..5708)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5691..5708)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(5762..5809)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(5833..6288)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5970..5991)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6182..6201
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(6343..7134)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7602..8190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8091..8110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8957..9315,1..256,327..458,662..977,
                     1039..1693,1768..2041,2443..2514))
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
ORIGIN
        1 aggagctggc gcaggatatt agatatccga aggacgttga cacattggcc accagagtga
       61 ccagcgccag gcagttgaag aagtgcagca ctccggcccg cagtccgatc atcggatagg
      121 caatcgccgt gaagaccagt ggcactgtga gaaaaagcgg caattcggca atcgttttgc
      181 ccagaaagta tgtgtcacag cgataaagtc gacttcgggc ctccctcata aaaactggca
      241 gctctgaggt gaacacctaa atcgaatcga ttcattagaa agttagtaaa ttattgaaat
      301 gcaaatgtat tctaaacatg acttacattt atcgtggcaa agacgttttg aaaggtcatg
      361 ttggtcagga agaggaagat ggctccgttg atattcatca cacccacttg cgtgagttgt
      421 tggcccaaaa agatgaggcc aatcaagatg gcaaccatct gcaaattaaa atgttactcg
      481 catctcatta atattcgcga gttaaatgaa atttatttat cttctgcaaa actataaact
      541 atacatctca ttgaaaaaaa ctaagaaggg tgtggaatca ggcaattcta tctaaaatct
      601 agcgaatttg tttccaagaa ttgtaagcgt tatatcattt gtttccactg gaaccactca
      661 ccgttgtctg aataagtcgc acttttacga ggagtggttc cttgagcacc gacagccagg
      721 atcgccacag gaccgcccgg aactgcatga accaggtggc cttgtaggtg tacccattct
      781 ccggctgctc cagtggcttc tccagatttt tggtggccaa caactgctcc atatcccggg
      841 ctactttgct aatggcaaaa ttgtcgcata tcttggcgat ccgatcacgg gactcgatct
      901 cccgtccggg cacaacggcc aacacctgta cgtaaaagtc cgccggattg tagttggtag
      961 gacactgggc acccacgctg gataggagtt gagatgttat gtaatactag atacccttaa
     1021 taaacacatc gaactcacta ggaaaagaag tcgacggctt cgctgggagt gcccaagaaa
     1081 gctaccctgc cctcggccat cagaaggatc ttgtcaaaga gctcaaacag ctcggaagac
     1141 ggctgatgaa tggtcaggat gacggtcttg cccttctgcg acagcttctt cagcacctgg
     1201 acgacgctgt gggcggtaaa ggagtccagt ccggaggtgg gctcatcgca gatcagaagc
     1261 ggcggatcgg ttagagcctc ggaggcgaat gccagacgct tcctttctcc gccggacaga
     1321 cctttcaccc tgccgggcac accgatgatc gtgtgctgac atttgctgag cgaaagctcc
     1381 tggatcacct gatccacgcg ggccactcgc tgccgatagg tcagatgtcg tggcatccgc
     1441 accatggctt ggaaaatcag gtgttccctg gccgttaggg agccgataaa gaggtcatcc
     1501 tgctggacat aggcgcacct ggcctgcatc tccttggcgt ccacaggttg gccattgagc
     1561 agtcgcatcc cggatggcga tacttggatg ccctgcggcg atcgaaaggc aagggcattc
     1621 agcagggtcg tctttccggc accggaactg cccatcacgg ccaaaagttc gcccggatag
     1681 gccacgccgc aaactgagtt tcaaattggt aattggaccc tttattaaga tttcacacag
     1741 atcagccgac tgcgaataga aactcaccgt tcttgagcaa atgtttcctg ggcgccggta
     1801 tgtgtcgctc gttgcagaat agtccgcgtg tccggttgac cagctgccgc catccggagc
     1861 ccggctgatt gaccgcccca aagatgtcca tattgtgcca ggcataggtg aggttctcgg
     1921 ctagttggcc gctccctgaa ccggagtcct ccggcggact gggtggccgg agcgtgccgt
     1981 agtttttggc ctgcccgaag ccctggttaa tgcagctctg cgaagccgct ccgctgtcac
     2041 cctgcaatga taggggatct caaatatcaa ctacaagcgt tatgctcatc taaccccgaa
     2101 caaaaagtac cccgaagtat cctacgaagt aggtttatac ttttatttat tttttgtgca
     2161 tctaggatca gcttaaaata tctggttgtt atattttttg taaaaaagaa tatagtcgaa
     2221 aatgaatgcc tttagatgtc ttgatcatga tatgatctca aaaattgtct tatatagcga
     2281 gaacagctac cagaataatc tgtttcgtgt cactatttgt ttgtgcaatt gcggtttggg
     2341 atttttgtgg gtcgcagttc tcacgccgca gacaatttga tgttgcaatc gcagttccta
     2401 tagatcaagt gaacttaaga tgtatgcaca tgtactactc acattgttca gatgctcggc
     2461 agatgggtgt ttgctgcctc cgcgaattaa tagctcctga tcctcttggc ccattgccgg
     2521 gatttttcac actttcccct gcttacccac ccaaaaccaa tcaccacccc aatcactcaa
     2581 aaaacaaaca aaaataagaa gcgagaggag ttttggcaca gcactttgtg tttaattgat
     2641 ggcgtaaacc gcttggagct tcgtcacgaa accgctgaca aaatgcaact gaaggcggac
     2701 attgacgcta cgtaacgcta caaacggtgg cgaaagagat agcggacgca gcggcgaaag
     2761 tgacggcgat atttctgtgg acagagaagg aggcaaacag cgctgacttt gagtggaatg
     2821 tcattttgag tgagaggtaa tcgaaagaac ctggtacatc aaataccctt ggatcgaagt
     2881 aaatttaaaa ctgatcagat aagttcaatg atatccagtg cagtaaaaaa aaaaaatgtt
     2941 ttttttatct actttccgca aaaatgggtt ttattaactt acatacatac tagaattcct
     3001 cctgacagcg aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3061 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3121 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3181 tcaatgtatc ttatcatgtc tggatcaagc ttacttgtac agctcatcca tgccgcccgt
     3241 gctatgacgt ccttcggcac gctcgtactg ctccacgatg gtgtagtcct cgttgtggct
     3301 ggtgatgtcc agcttgatgt tcacgttgta ggcgccgggc aattgcacgg gcttcttggc
     3361 cttgtaggtg gtcttcacct cggcgtcgta gtggcctcca tccttcagct tcaggcgctg
     3421 cttgatctcg cccttcagag cgccgtcctc gggatacatg cgctcgctgg aagcctccca
     3481 gcccatggtc ttcttctgca tcaccgggcc atcggaggga aagttcgtgc cgcgcagctt
     3541 gaccttgtag atgaactcgc cgtcctgcaa gctggagtcc tgggtcacgg tcaccactcc
     3601 gccgtcctcg aagttcatca cgcgctccca cttgaagcct tcggggaagc tcagcttcag
     3661 gtagtcggga atgtcggccg ggtgcttcac gtaggccttg gagccgtaca tgaactgggg
     3721 gctcaagatg tcccaggcaa agggcaacgg gcctcccttg gtcaccttca gtttggcggt
     3781 ctgggtgcct tcgtagggac gtccctcgcc ttcgccctcg atctcgaact cgtggccatt
     3841 cacgctgccc tccatgtgga ccttgaagcg catgaactcc ttgatgatgg ccatgttatc
     3901 ctcttctccc ttggacacca tttggtttgg tatgcgtctt gtgattcaaa gttggcttat
     3961 tcaaaggata ttaacgaagg cagcggcacg tctgctcggc tcgaacattc attcacaact
     4021 gatgctctca gccaccccgc gcccttttat accgctgcgc tcgatccccg ggcgagctcg
     4081 ggaaaggata agcgattacc cgaggataag cgattacccg gataagcgat tacccgtgag
     4141 gataattgtt tatccgagga taagcgatta cccctcctga cagcggaaca aacgacccaa
     4201 cacccgtgcg ttttattctg tctttttatt gccaagctta gttacgggta tacttcaggg
     4261 gaatcagttc ctcaatggtg gtcttgacca gcttgccgtt catctcaatc agcacgaagc
     4321 aatcgggggc gtaatcgcta atcagctcac ggcacattcc acacggggac acgacacgga
     4381 tgctacggtc cacctcatcg gagtatgggt gacgcacggc cacaatggta tcaaaatcct
     4441 tctggccgtt gctcacggca ctgccaatgg caatggcctc ggcacacacg gtcacgcgtc
     4501 cgatgtaggc ctcaatgtgc acggcactga taatctcgcc cgtcttggtg cgaatggcag
     4561 ctcccacgtg gtgcttgtta tcctcgtaga gcatcgtaat cttctcggta gccacctcca
     4621 ccagttccag gtcctgctgg ctgatgttga aggttttcat tttgttttcc tccttatgtt
     4681 aagcttactc agttattata tcataaatat ctgtgtcaag aataaactcc cacatggatt
     4741 cgagtattga gtgttctctt cttgtattca ataattactt cttggcagat ttcagtagtt
     4801 gcagttgatt tacttggttg ctggttactt ttaattgatt cactttaact tgcactttac
     4861 tgcagattgt ttagcttgtt cagctgcgct tgtttatttg cttagctttc gcttagcgac
     4921 gtgttcactt tgcttgtttg aattgaattg tcgctccgta gacgaagcgc ctctatttat
     4981 actccggcgc tcttttcgcg aacattcgag gcgcgctctc tcgaaccaac gagagcagta
     5041 tgccgtttac tgtgtgacag agtgagagag cattagtgca gagagggagc gacccaaaaa
     5101 gaaaagagag aataacgaat aacggccaga gaaatttctc gagttttctt tctgccaaac
     5161 aaatgaccta ccacaataac cagtttgttt tgggattcta gctcctgaca gcgcatcatg
     5221 atggaccaga tgggtgaggt ggagtacgcg cccggggagc ccaagggcac gccctggcac
     5281 ccgcaccgcg gcttcgagac agtgacctac atcgtcgacc aattcctccc tgggatccgg
     5341 agagagaccc tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5401 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgtcattcgc cattcaggct
     5461 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5521 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5581 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tcgagctcgg
     5641 tacccgggga tcctctagag tcgacctgca ggcatgcaag cttgagtatt ctatagtgtc
     5701 acctaaatag cttggcgtaa tcatggtcat gatctgcctc ctgggtttag ttcctcacct
     5761 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca ggggtctctc
     5821 gctgatccgt taaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5881 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5941 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6001 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgctggc ccactacgtg
     6061 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6121 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6181 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6241 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa catgaagttt
     6301 taaatcaatc taaagtatat atgtgtaaca ttggtctagt gattatttgc cgactacctt
     6361 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     6421 acgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     6481 ggccggcagg cgctccattg cccagtcggc agctacatcc ttcggcgcga ttttgccggt
     6541 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     6601 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     6661 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     6721 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     6781 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     6841 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     6901 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     6961 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     7021 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     7081 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcataatgtt
     7141 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
     7201 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc
     7261 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
     7321 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gcttacgaac
     7381 cgaacaggct tatgtccact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc
     7441 ggcaaccttg ggtagcagcg aagtcgaggc atttctgtcc tggctggtca caaaaggatc
     7501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8281 gataaccgtg tcaagcgatg catctacaca aggaacaaac actggatgtc actttcagtt
     8341 caaattgtaa cgctaatcac tccgaacagg tcacaaaaaa ttaccttaaa aagtcataat
     8401 attaaattag aataaatata gctgtgaggg aaatatatac aaatatattg gagcaaataa
     8461 attgtacata caaatattta ttactaattt ctattgagcc gaaatgaacc actcggaacc
     8521 atttgagcga accgaatcgc gcggaactaa cgacagtcgc tccaaggtcg tcgaacaaaa
     8581 ggtgaatgtg ttgcggagag cgggtgggag acagcgaaag agcaactacg aaacgtggtg
     8641 tggtggaggt gaattatgaa gagggcgcgc gatttgaaaa gtatgtatat aaaaaatata
     8701 tcccggtgtt ttatgtagcg ataaacgagt ttttgatgta aggtatgcag gtgtgtaagt
     8761 cttttggtta gaagacaaat ccaaagtcta cttgtgggga tgttcgaagg ggaaatactt
     8821 gtattctata ggtcatatct tgtttttatt ggcacaaata taattacatt agctttttga
     8881 gggggcaata aacagtaaac acgatggtaa taatggtaaa aaaaaaaaac aagcagttat
     8941 ttcggatata tgtcggctac tccttgcgtc gggcccgaag tcttagagcc agatatgcga
     9001 gcacccggaa gctcacgatg agaatggcca gacccacgta gtccagcggc agatcggcgg
     9061 cggagaagtt aagtgtctcc aggatgacct tgcccgaact ggggcacgtg gtgttcgacg
     9121 atgtgcagct aatttcgccc ggctccacgt ccgcccattg gttaatcagc agaccctcgt
     9181 tggcgtaacg gaaccatgag aggtacgaca accatttgag gtatactggc accgagcccg
     9241 agttcaagaa gaagccgcca aagagcagga atggtatgat aaccggcgga cccacagaca
     9301 gcgccatcga ggtcg
//
