LOCUS       sequence-326310-        9318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xUAS 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 9318)
  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 9318)
  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-5xUAS-mCherry-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(74..688,759..890,1094..1409,1471..2125,
                     2200..2473,2875..2946))
                     /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"
     primer_bind     3398..3416
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     polyA_signal    3504..3638
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3541..3560)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3595..3614
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5216..5237)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    5664..5733
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     5790..5809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6003..6025
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6017..6034
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6018..6034
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6041..6060
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6041..6059
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6062..6118
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(6125..6143)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6126..6143)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(6197..6244)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(6268..6723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6405..6426)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6617..6636
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(6778..7569)
                     /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      8037..8625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8526..8545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcaataaac agtaaacacg atggtaataa tggtaaaaaa aaaaaacaag cagttatttc
       61 ggatatatgt cggctactcc ttgcgtcggg cccgaagtct tagagccaga tatgcgagca
      121 cccggaagct cacgatgaga atggccagac ccacgtagtc cagcggcaga tcggcggcgg
      181 agaagttaag tgtctccagg atgaccttgc ccgaactggg gcacgtggtg ttcgacgatg
      241 tgcagctaat ttcgcccggc tccacgtccg cccattggtt aatcagcaga ccctcgttgg
      301 cgtaacggaa ccatgagagg tacgacaacc atttgaggta tactggcacc gagcccgagt
      361 tcaagaagaa gccgccaaag agcaggaatg gtatgataac cggcggaccc acagacagcg
      421 ccatcgaggt cgaggagctg gcgcaggata ttagatatcc gaaggacgtt gacacattgg
      481 ccaccagagt gaccagcgcc aggcagttga agaagtgcag cactccggcc cgcagtccga
      541 tcatcggata ggcaatcgcc gtgaagacca gtggcactgt gagaaaaagc ggcaattcgg
      601 caatcgtttt gcccagaaag tatgtgtcac agcgataaag tcgacttcgg gcctccctca
      661 taaaaactgg cagctctgag gtgaacacct aaatcgaatc gattcattag aaagttagta
      721 aattattgaa atgcaaatgt attctaaaca tgacttacat ttatcgtggc aaagacgttt
      781 tgaaaggtca tgttggtcag gaagaggaag atggctccgt tgatattcat cacacccact
      841 tgcgtgagtt gttggcccaa aaagatgagg ccaatcaaga tggcaaccat ctgcaaatta
      901 aaatgttact cgcatctcat taatattcgc gagttaaatg aaatttattt atcttctgca
      961 aaactataaa ctatacatct cattgaaaaa aactaagaag ggtgtggaat caggcaattc
     1021 tatctaaaat ctagcgaatt tgtttccaag aattgtaagc gttatatcat ttgtttccac
     1081 tggaaccact caccgttgtc tgaataagtc gcacttttac gaggagtggt tccttgagca
     1141 ccgacagcca ggatcgccac aggaccgccc ggaactgcat gaaccaggtg gccttgtagg
     1201 tgtacccatt ctccggctgc tccagtggct tctccagatt tttggtggcc aacaactgct
     1261 ccatatcccg ggctactttg ctaatggcaa aattgtcgca tatcttggcg atccgatcac
     1321 gggactcgat ctcccgtccg ggcacaacgg ccaacacctg tacgtaaaag tccgccggat
     1381 tgtagttggt aggacactgg gcacccacgc tggataggag ttgagatgtt atgtaatact
     1441 agataccctt aataaacaca tcgaactcac taggaaaaga agtcgacggc ttcgctggga
     1501 gtgcccaaga aagctaccct gccctcggcc atcagaagga tcttgtcaaa gagctcaaac
     1561 agctcggaag acggctgatg aatggtcagg atgacggtct tgcccttctg cgacagcttc
     1621 ttcagcacct ggacgacgct gtgggcggta aaggagtcca gtccggaggt gggctcatcg
     1681 cagatcagaa gcggcggatc ggttagagcc tcggaggcga atgccagacg cttcctttct
     1741 ccgccggaca gacctttcac cctgccgggc acaccgatga tcgtgtgctg acatttgctg
     1801 agcgaaagct cctggatcac ctgatccacg cgggccactc gctgccgata ggtcagatgt
     1861 cgtggcatcc gcaccatggc ttggaaaatc aggtgttccc tggccgttag ggagccgata
     1921 aagaggtcat cctgctggac ataggcgcac ctggcctgca tctccttggc gtccacaggt
     1981 tggccattga gcagtcgcat cccggatggc gatacttgga tgccctgcgg cgatcgaaag
     2041 gcaagggcat tcagcagggt cgtctttccg gcaccggaac tgcccatcac ggccaaaagt
     2101 tcgcccggat aggccacgcc gcaaactgag tttcaaattg gtaattggac cctttattaa
     2161 gatttcacac agatcagccg actgcgaata gaaactcacc gttcttgagc aaatgtttcc
     2221 tgggcgccgg tatgtgtcgc tcgttgcaga atagtccgcg tgtccggttg accagctgcc
     2281 gccatccgga gcccggctga ttgaccgccc caaagatgtc catattgtgc caggcatagg
     2341 tgaggttctc ggctagttgg ccgctccctg aaccggagtc ctccggcgga ctgggtggcc
     2401 ggagcgtgcc gtagtttttg gcctgcccga agccctggtt aatgcagctc tgcgaagccg
     2461 ctccgctgtc accctgcaat gataggggat ctcaaatatc aactacaagc gttatgctca
     2521 tctaaccccg aacaaaaagt accccgaagt atcctacgaa gtaggtttat acttttattt
     2581 attttttgtg catctaggat cagcttaaaa tatctggttg ttatattttt tgtaaaaaag
     2641 aatatagtcg aaaatgaatg cctttagatg tcttgatcat gatatgatct caaaaattgt
     2701 cttatatagc gagaacagct accagaataa tctgtttcgt gtcactattt gtttgtgcaa
     2761 ttgcggtttg ggatttttgt gggtcgcagt tctcacgccg cagacaattt gatgttgcaa
     2821 tcgcagttcc tatagatcaa gtgaacttaa gatgtatgca catgtactac tcacattgtt
     2881 cagatgctcg gcagatgggt gtttgctgcc tccgcgaatt aatagctcct gatcctcttg
     2941 gcccattgcc gggatttttc acactttccc ctgcttaccc acccaaaacc aatcaccacc
     3001 ccaatcactc aaaaaacaaa caaaaataag aagcgagagg agttttggca cagcactttg
     3061 tgtttaattg atggcgtaaa ccgcttggag cttcgtcacg aaaccgctga caaaatgcaa
     3121 ctgaaggcgg acattgacgc tacgtaacgc tacaaacggt ggcgaaagag atagcggacg
     3181 cagcggcgaa agtgacggcg atatttctgt ggacagagaa ggaggcaaac agcgctgact
     3241 ttgagtggaa tgtcattttg agtgagaggt aatcgaaaga acctggtaca tcaaataccc
     3301 ttggatcgaa gtaaatttaa aactgatcag ataagttcaa tgatatccag tgcagtaaaa
     3361 aaaaaaaatg ttttttttat ctactttccg caaaaatggg ttttattaac ttacatacat
     3421 actagaattc ctcctgacag cgaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3481 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3541 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3601 tgtccaaact catcaatgta tcttatcatg tctggatcaa gcttacttgt acagctcatc
     3661 catgccgccc gtgctatgac gtccttcggc acgctcgtac tgctccacga tggtgtagtc
     3721 ctcgttgtgg ctggtgatgt ccagcttgat gttcacgttg taggcgccgg gcaattgcac
     3781 gggcttcttg gccttgtagg tggtcttcac ctcggcgtcg tagtggcctc catccttcag
     3841 cttcaggcgc tgcttgatct cgcccttcag agcgccgtcc tcgggataca tgcgctcgct
     3901 ggaagcctcc cagcccatgg tcttcttctg catcaccggg ccatcggagg gaaagttcgt
     3961 gccgcgcagc ttgaccttgt agatgaactc gccgtcctgc aagctggagt cctgggtcac
     4021 ggtcaccact ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cttcggggaa
     4081 gctcagcttc aggtagtcgg gaatgtcggc cgggtgcttc acgtaggcct tggagccgta
     4141 catgaactgg gggctcaaga tgtcccaggc aaagggcaac gggcctccct tggtcacctt
     4201 cagtttggcg gtctgggtgc cttcgtaggg acgtccctcg ccttcgccct cgatctcgaa
     4261 ctcgtggcca ttcacgctgc cctccatgtg gaccttgaag cgcatgaact ccttgatgat
     4321 ggccatgtta tcctcttctc ccttggacac catttggttt ggtatgcgtc ttgtgattca
     4381 aagttggctt attcaaagga tattaacgaa ggcagcggca cgtctgctcg gctcgaacat
     4441 tcattcacaa ctgatgctct cagccacccc gcgccctttt ataccgctgc gctcgatccc
     4501 cgggcgagct cgggacggag gacagtactc cgctcggagg acagtactcc gctcggagga
     4561 cagtactccg ctcggaggac agtactccgc tcggaggaca gtactccgct cctgacagcg
     4621 gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt ttattgccaa gcttagttac
     4681 gggtatactt caggggaatc agttcctcaa tggtggtctt gaccagcttg ccgttcatct
     4741 caatcagcac gaagcaatcg ggggcgtaat cgctaatcag ctcacggcac attccacacg
     4801 gggacacgac acggatgcta cggtccacct catcggagta tgggtgacgc acggccacaa
     4861 tggtatcaaa atccttctgg ccgttgctca cggcactgcc aatggcaatg gcctcggcac
     4921 acacggtcac gcgtccgatg taggcctcaa tgtgcacggc actgataatc tcgcccgtct
     4981 tggtgcgaat ggcagctccc acgtggtgct tgttatcctc gtagagcatc gtaatcttct
     5041 cggtagccac ctccaccagt tccaggtcct gctggctgat gttgaaggtt ttcattttgt
     5101 tttcctcctt atgttaagct tactcagtta ttatatcata aatatctgtg tcaagaataa
     5161 actcccacat ggattcgagt attgagtgtt ctcttcttgt attcaataat tacttcttgg
     5221 cagatttcag tagttgcagt tgatttactt ggttgctggt tacttttaat tgattcactt
     5281 taacttgcac tttactgcag attgtttagc ttgttcagct gcgcttgttt atttgcttag
     5341 ctttcgctta gcgacgtgtt cactttgctt gtttgaattg aattgtcgct ccgtagacga
     5401 agcgcctcta tttatactcc ggcgctcttt tcgcgaacat tcgaggcgcg ctctctcgaa
     5461 ccaacgagag cagtatgccg tttactgtgt gacagagtga gagagcatta gtgcagagag
     5521 ggagcgaccc aaaaagaaaa gagagaataa cgaataacgg ccagagaaat ttctcgagtt
     5581 ttctttctgc caaacaaatg acctaccaca ataaccagtt tgttttggga ttctagctcc
     5641 tgacagcgca tcatgatgga ccagatgggt gaggtggagt acgcgcccgg ggagcccaag
     5701 ggcacgccct ggcacccgca ccgcggcttc gagacagtga cctacatcgt cgaccaattc
     5761 ctccctggga tccggagaga gaccctatgc ggcatcagag cagattgtac tgagagtgca
     5821 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgtca
     5881 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5941 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6001 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     6061 cgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttga
     6121 gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatgatct gcctcctggg
     6181 tttagttcct caccttgtcg tattatacta tgccgatata ctatgccgat gattaattgt
     6241 caacaggggt ctctcgctga tccgttaaaa ttgtaagcgt taatattttg ttaaaattcg
     6301 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6361 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6421 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6481 ctggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6541 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6601 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6661 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6721 cgtaacatga agttttaaat caatctaaag tatatatgtg taacattggt ctagtgatta
     6781 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     6841 tgcgcgcgag gccaaacgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     6901 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcta catccttcgg
     6961 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     7021 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     7081 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     7141 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     7201 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     7261 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     7321 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     7381 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     7441 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     7501 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     7561 ttccctcata atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     7621 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     7681 catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     7741 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     7801 ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca
     7861 cggtgtgcgt cacccggcaa ccttgggtag cagcgaagtc gaggcatttc tgtcctggct
     7921 ggtcacaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7981 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8041 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8101 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8161 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     8221 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8281 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8341 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8401 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8461 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8521 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8581 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8641 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8701 tcccctgatt ctgtggataa ccgtgtcaag cgatgcatct acacaaggaa caaacactgg
     8761 atgtcacttt cagttcaaat tgtaacgcta atcactccga acaggtcaca aaaaattacc
     8821 ttaaaaagtc ataatattaa attagaataa atatagctgt gagggaaata tatacaaata
     8881 tattggagca aataaattgt acatacaaat atttattact aatttctatt gagccgaaat
     8941 gaaccactcg gaaccatttg agcgaaccga atcgcgcgga actaacgaca gtcgctccaa
     9001 ggtcgtcgaa caaaaggtga atgtgttgcg gagagcgggt gggagacagc gaaagagcaa
     9061 ctacgaaacg tggtgtggtg gaggtgaatt atgaagaggg cgcgcgattt gaaaagtatg
     9121 tatataaaaa atatatcccg gtgttttatg tagcgataaa cgagtttttg atgtaaggta
     9181 tgcaggtgtg taagtctttt ggttagaaga caaatccaaa gtctacttgt ggggatgttc
     9241 gaaggggaaa tacttgtatt ctataggtca tatcttgttt ttattggcac aaatataatt
     9301 acattagctt tttgaggg
//
