LOCUS       sequence-326303-        9324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xQUAS EBFP2 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 9324)
  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 9324)
  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-EBFP2-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(950..1538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1030..1049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2006..2797
                     /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      2852..3307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2939..2958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3149..3170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3331..3378
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        3432..3450
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3432..3449
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    3457..3513
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3515..3534)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3516..3534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3541..3558)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3541..3557)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3550..3572)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3766..3785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3842..3911
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     4338..4359
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     polyA_signal    5943..6077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5967..5986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6021..6040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6165..6183)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(6635..6706,7108..7381,7456..8110,8172..8487,8691..8822,
                     8893..9324,1..183)
                     /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 ggcgaaatta gctgcacatc gtcgaacacc acgtgcccca gttcgggcaa ggtcatcctg
       61 gagacactta acttctccgc cgccgatctg ccgctggact acgtgggtct ggccattctc
      121 atcgtgagct tccgggtgct cgcatatctg gctctaagac ttcgggcccg acgcaaggag
      181 tagccgacat atatccgaaa taactgcttg tttttttttt ttaccattat taccatcgtg
      241 tttactgttt attgccccct caaaaagcta atgtaattat atttgtgcca ataaaaacaa
      301 gatatgacct atagaataca agtatttccc cttcgaacat ccccacaagt agactttgga
      361 tttgtcttct aaccaaaaga cttacacacc tgcatacctt acatcaaaaa ctcgtttatc
      421 gctacataaa acaccgggat atatttttta tatacatact tttcaaatcg cgcgccctct
      481 tcataattca cctccaccac accacgtttc gtagttgctc tttcgctgtc tcccacccgc
      541 tctccgcaac acattcacct tttgttcgac gaccttggag cgactgtcgt tagttccgcg
      601 cgattcggtt cgctcaaatg gttccgagtg gttcatttcg gctcaataga aattagtaat
      661 aaatatttgt atgtacaatt tatttgctcc aatatatttg tatatatttc cctcacagct
      721 atatttattc taatttaata ttatgacttt ttaaggtaat tttttgtgac ctgttcggag
      781 tgattagcgt tacaatttga actgaaagtg acatccagtg tttgttcctt gtgtagatgc
      841 atcgcttgac acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      901 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1141 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1381 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1621 caaaaaggat cttcacctag atccttttgt gaccagccag gacagaaatg cctcgacttc
     1681 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     1741 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     1801 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     1861 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1921 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     1981 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     2041 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     2101 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     2161 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     2221 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     2281 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     2341 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     2401 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     2461 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     2521 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     2581 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtagctg ccgactgggc
     2641 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     2701 tggacaagaa gaagatcgtt tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     2761 cgtgaaaggc gagatcacca aggtagtcgg caaataatca ctagaccaat gttacacata
     2821 tatactttag attgatttaa aacttcatgt tacgcgccct gtagcggcgc attaagcgcg
     2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3061 cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt ttttcgccct
     3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3181 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3301 acaattttaa cggatcagcg agagacccct gttgacaatt aatcatcggc atagtatatc
     3361 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgaccatgat
     3421 tacgccaagc tatttaggtg acactataga atactcaagc ttgcatgcct gcaggtcgac
     3481 tctagaggat ccccgggtac cgagctcgaa ttcgccctat agtgagtcgt attacaattc
     3541 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3601 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3661 cccttcccaa cagttgcgca gcctgaatgg cgaatgacgc ctgatgcggt attttctcct
     3721 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3781 tgccgcatag ggtctctctc cggatcccag ggaggaattg gtcgacgatg taggtcactg
     3841 tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
     3901 cctcacccat ctggtccatc atgatgcgct gtcaggagct agaatcccaa aacaaactgg
     3961 ttattgtggt aggtcatttg tttggcagaa agaaaactcg agaaatttct ctggccgtta
     4021 ttcgttattc tctcttttct ttttgggtcg ctccctctct gcactaatgc tctctcactc
     4081 tgtcacacag taaacggcat actgctctcg ttggttcgag agagcgcgcc tcgaatgttc
     4141 gcgaaaagag cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc
     4201 aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga
     4261 acaagctaaa caatctgcag taaagtgcaa gttaaagtga atcaattaaa agtaaccagc
     4321 aaccaagtaa atcaactgca actactgaaa tctgccaaga agtaattatt gaatacaaga
     4381 agagaacact caatactcga atccatgtgg gagtttattc ttgacacaga tatttatgat
     4441 ataataactg agtaagctta acataaggag gaaaacaaaa tgaaaacctt caacatcagc
     4501 cagcaggacc tggaactggt ggaggtggct accgagaaga ttacgatgct ctacgaggat
     4561 aacaagcacc acgtgggagc tgccattcgc accaagacgg gcgagattat cagtgccgtg
     4621 cacattgagg cctacatcgg acgcgtgacc gtgtgtgccg aggccattgc cattggcagt
     4681 gccgtgagca acggccagaa ggattttgat accattgtgg ccgtgcgtca cccatactcc
     4741 gatgaggtgg accgtagcat ccgtgtcgtg tccccgtgtg gaatgtgccg tgagctgatt
     4801 agcgattacg cccccgattg cttcgtgctg attgagatga acggcaagct ggtcaagacc
     4861 accattgagg aactgattcc cctgaagtat acccgtaact aagcttggca ataaaaagac
     4921 agaataaaac gcacgggtgt tgggtcgttt gttccgctgt caggaggggt aatcgcttat
     4981 cctcggataa acaattatcc tcacgggtaa tcgcttatcc gggtaatcgc ttatcctcgg
     5041 gtaatcgctt atcctttccc gagctcgccc ggggatcgag cgcagcggta taaaagggcg
     5101 cggggtggct gagagcatca gttgtgaatg aatgttcgag ccgagcagac gtgccgctgc
     5161 cttcgttaat atcctttgaa taagccaact ttgaatcaca agacgcatac caaaccaaat
     5221 ggtgtccaag ggcgaggagc tgttcaccgg cgtggtgccc atcctggtgg agctggatgg
     5281 cgacgtgaac ggccacaagt tcagcgtgcg cggcgagggc gagggcgacg ccaccaacgg
     5341 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     5401 ggtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc taccccgatc acatgaagca
     5461 gcacgatttc ttcaagagcg ccatgcccga gggctacgtg caggagcgca ccattttctt
     5521 caatgacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg ataccctggt
     5581 gaaccgcatc gagctgaagg gcgtggattt caaggaggat ggcaacatcc tgggccacaa
     5641 gctggagtac aacttcaaca gccacaacat ctacatcatg gccgtgaagc agaagaacgg
     5701 catcaaggtc aacttcaaga tccgccacaa cgtggaggat ggctcagtgc agctggccga
     5761 tcactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg atagccacta
     5821 cctgagcacc cagagcgtgc tgtccaagga tcccaacgag aagcgcgatc acatggtgct
     5881 gctggagttc cgcaccgccg ccggcatcac cctgggcatg gatgagctgt acaagtaagc
     5941 ttgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     6001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     6061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     6121 aggtgtggga ggttttttcg ctgtcaggag gaattctagt atgtatgtaa gttaataaaa
     6181 cccatttttg cggaaagtag ataaaaaaaa catttttttt ttttactgca ctggatatca
     6241 ttgaacttat ctgatcagtt ttaaatttac ttcgatccaa gggtatttga tgtaccaggt
     6301 tctttcgatt acctctcact caaaatgaca ttccactcaa agtcagcgct gtttgcctcc
     6361 ttctctgtcc acagaaatat cgccgtcact ttcgccgctg cgtccgctat ctctttcgcc
     6421 accgtttgta gcgttacgta gcgtcaatgt ccgccttcag ttgcattttg tcagcggttt
     6481 cgtgacgaag ctccaagcgg tttacgccat caattaaaca caaagtgctg tgccaaaact
     6541 cctctcgctt cttatttttg tttgtttttt gagtgattgg ggtggtgatt ggttttgggt
     6601 gggtaagcag gggaaagtgt gaaaaatccc ggcaatgggc caagaggatc aggagctatt
     6661 aattcgcgga ggcagcaaac acccatctgc cgagcatctg aacaatgtga gtagtacatg
     6721 tgcatacatc ttaagttcac ttgatctata ggaactgcga ttgcaacatc aaattgtctg
     6781 cggcgtgaga actgcgaccc acaaaaatcc caaaccgcaa ttgcacaaac aaatagtgac
     6841 acgaaacaga ttattctggt agctgttctc gctatataag acaatttttg agatcatatc
     6901 atgatcaaga catctaaagg cattcatttt cgactatatt cttttttaca aaaaatataa
     6961 caaccagata ttttaagctg atcctagatg cacaaaaaat aaataaaagt ataaacctac
     7021 ttcgtaggat acttcggggt actttttgtt cggggttaga tgagcataac gcttgtagtt
     7081 gatatttgag atcccctatc attgcagggt gacagcggag cggcttcgca gagctgcatt
     7141 aaccagggct tcgggcaggc caaaaactac ggcacgctcc ggccacccag tccgccggag
     7201 gactccggtt cagggagcgg ccaactagcc gagaacctca cctatgcctg gcacaatatg
     7261 gacatctttg gggcggtcaa tcagccgggc tccggatggc ggcagctggt caaccggaca
     7321 cgcggactat tctgcaacga gcgacacata ccggcgccca ggaaacattt gctcaagaac
     7381 ggtgagtttc tattcgcagt cggctgatct gtgtgaaatc ttaataaagg gtccaattac
     7441 caatttgaaa ctcagtttgc ggcgtggcct atccgggcga acttttggcc gtgatgggca
     7501 gttccggtgc cggaaagacg accctgctga atgcccttgc ctttcgatcg ccgcagggca
     7561 tccaagtatc gccatccggg atgcgactgc tcaatggcca acctgtggac gccaaggaga
     7621 tgcaggccag gtgcgcctat gtccagcagg atgacctctt tatcggctcc ctaacggcca
     7681 gggaacacct gattttccaa gccatggtgc ggatgccacg acatctgacc tatcggcagc
     7741 gagtggcccg cgtggatcag gtgatccagg agctttcgct cagcaaatgt cagcacacga
     7801 tcatcggtgt gcccggcagg gtgaaaggtc tgtccggcgg agaaaggaag cgtctggcat
     7861 tcgcctccga ggctctaacc gatccgccgc ttctgatctg cgatgagccc acctccggac
     7921 tggactcctt taccgcccac agcgtcgtcc aggtgctgaa gaagctgtcg cagaagggca
     7981 agaccgtcat cctgaccatt catcagccgt cttccgagct gtttgagctc tttgacaaga
     8041 tccttctgat ggccgagggc agggtagctt tcttgggcac tcccagcgaa gccgtcgact
     8101 tcttttccta gtgagttcga tgtgtttatt aagggtatct agtattacat aacatctcaa
     8161 ctcctatcca gcgtgggtgc ccagtgtcct accaactaca atccggcgga cttttacgta
     8221 caggtgttgg ccgttgtgcc cggacgggag atcgagtccc gtgatcggat cgccaagata
     8281 tgcgacaatt ttgccattag caaagtagcc cgggatatgg agcagttgtt ggccaccaaa
     8341 aatctggaga agccactgga gcagccggag aatgggtaca cctacaaggc cacctggttc
     8401 atgcagttcc gggcggtcct gtggcgatcc tggctgtcgg tgctcaagga accactcctc
     8461 gtaaaagtgc gacttattca gacaacggtg agtggttcca gtggaaacaa atgatataac
     8521 gcttacaatt cttggaaaca aattcgctag attttagata gaattgcctg attccacacc
     8581 cttcttagtt tttttcaatg agatgtatag tttatagttt tgcagaagat aaataaattt
     8641 catttaactc gcgaatatta atgagatgcg agtaacattt taatttgcag atggttgcca
     8701 tcttgattgg cctcatcttt ttgggccaac aactcacgca agtgggtgtg atgaatatca
     8761 acggagccat cttcctcttc ctgaccaaca tgacctttca aaacgtcttt gccacgataa
     8821 atgtaagtca tgtttagaat acatttgcat ttcaataatt tactaacttt ctaatgaatc
     8881 gattcgattt aggtgttcac ctcagagctg ccagttttta tgagggaggc ccgaagtcga
     8941 ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattgcc gctttttctc
     9001 acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg ggccggagtg
     9061 ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc aacgtccttc
     9121 ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt gggtccgccg
     9181 gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc ggtgccagta
     9241 tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg tctgctgatt
     9301 aaccaatggg cggacgtgga gccg
//
