LOCUS       sequence-326318-        9324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xQUAS sfGFP 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-sfGFP-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(973..1561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1053..1072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2029..2820
                     /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      2875..3330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2962..2981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3172..3193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3354..3401
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        3455..3473
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3455..3472
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    3480..3536
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3538..3557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3539..3557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3564..3581)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3564..3580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3573..3595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3789..3808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    3865..3934
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     4361..4382
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     polyA_signal    5966..6100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5990..6009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6044..6063
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6188..6206)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(6658..6729,7131..7404,7479..8133,8195..8510,8714..8845,
                     8916..9324,1..206)
                     /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 accaatgggc ggacgtggag ccgggcgaaa ttagctgcac atcgtcgaac accacgtgcc
       61 ccagttcggg caaggtcatc ctggagacac ttaacttctc cgccgccgat ctgccgctgg
      121 actacgtggg tctggccatt ctcatcgtga gcttccgggt gctcgcatat ctggctctaa
      181 gacttcgggc ccgacgcaag gagtagccga catatatccg aaataactgc ttgttttttt
      241 tttttaccat tattaccatc gtgtttactg tttattgccc cctcaaaaag ctaatgtaat
      301 tatatttgtg ccaataaaaa caagatatga cctatagaat acaagtattt ccccttcgaa
      361 catccccaca agtagacttt ggatttgtct tctaaccaaa agacttacac acctgcatac
      421 cttacatcaa aaactcgttt atcgctacat aaaacaccgg gatatatttt ttatatacat
      481 acttttcaaa tcgcgcgccc tcttcataat tcacctccac cacaccacgt ttcgtagttg
      541 ctctttcgct gtctcccacc cgctctccgc aacacattca ccttttgttc gacgaccttg
      601 gagcgactgt cgttagttcc gcgcgattcg gttcgctcaa atggttccga gtggttcatt
      661 tcggctcaat agaaattagt aataaatatt tgtatgtaca atttatttgc tccaatatat
      721 ttgtatatat ttccctcaca gctatattta ttctaattta atattatgac tttttaaggt
      781 aattttttgt gacctgttcg gagtgattag cgttacaatt tgaactgaaa gtgacatcca
      841 gtgtttgttc cttgtgtaga tgcatcgctt gacacggtta tccacagaat caggggataa
      901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1381 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgtgaccagc
     1681 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     1741 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     1801 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     1861 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     1921 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     1981 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     2041 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     2101 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     2161 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     2221 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     2281 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     2341 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     2401 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     2461 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     2521 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     2581 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     2641 aaggatgtag ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     2701 cttgaagcta gacaggctta tcttggacaa gaagaagatc gtttggcctc gcgcgcagat
     2761 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     2821 tcactagacc aatgttacac atatatactt tagattgatt taaaacttca tgttacgcgc
     2881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccagcgc
     3121 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3181 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3241 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3301 attttaacaa aatattaacg cttacaattt taacggatca gcgagagacc cctgttgaca
     3361 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     3421 aggaggcaga tcatgaccat gattacgcca agctatttag gtgacactat agaatactca
     3481 agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc gaattcgccc
     3541 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3601 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3661 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatga
     3721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3781 actctcagta caatctgctc tgatgccgca tagggtctct ctccggatcc cagggaggaa
     3841 ttggtcgacg atgtaggtca ctgtctcgaa gccgcggtgc gggtgccagg gcgtgccctt
     3901 gggctccccg ggcgcgtact ccacctcacc catctggtcc atcatgatgc gctgtcagga
     3961 gctagaatcc caaaacaaac tggttattgt ggtaggtcat ttgtttggca gaaagaaaac
     4021 tcgagaaatt tctctggccg ttattcgtta ttctctcttt tctttttggg tcgctccctc
     4081 tctgcactaa tgctctctca ctctgtcaca cagtaaacgg catactgctc tcgttggttc
     4141 gagagagcgc gcctcgaatg ttcgcgaaaa gagcgccgga gtataaatag aggcgcttcg
     4201 tctacggagc gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta
     4261 agcaaataaa caagcgcagc tgaacaagct aaacaatctg cagtaaagtg caagttaaag
     4321 tgaatcaatt aaaagtaacc agcaaccaag taaatcaact gcaactactg aaatctgcca
     4381 agaagtaatt attgaataca agaagagaac actcaatact cgaatccatg tgggagttta
     4441 ttcttgacac agatatttat gatataataa ctgagtaagc ttaacataag gaggaaaaca
     4501 aaatgaaaac cttcaacatc agccagcagg acctggaact ggtggaggtg gctaccgaga
     4561 agattacgat gctctacgag gataacaagc accacgtggg agctgccatt cgcaccaaga
     4621 cgggcgagat tatcagtgcc gtgcacattg aggcctacat cggacgcgtg accgtgtgtg
     4681 ccgaggccat tgccattggc agtgccgtga gcaacggcca gaaggatttt gataccattg
     4741 tggccgtgcg tcacccatac tccgatgagg tggaccgtag catccgtgtc gtgtccccgt
     4801 gtggaatgtg ccgtgagctg attagcgatt acgcccccga ttgcttcgtg ctgattgaga
     4861 tgaacggcaa gctggtcaag accaccattg aggaactgat tcccctgaag tatacccgta
     4921 actaagcttg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttccgc
     4981 tgtcaggagg ggtaatcgct tatcctcgga taaacaatta tcctcacggg taatcgctta
     5041 tccgggtaat cgcttatcct cgggtaatcg cttatccttt cccgagctcg cccggggatc
     5101 gagcgcagcg gtataaaagg gcgcggggtg gctgagagca tcagttgtga atgaatgttc
     5161 gagccgagca gacgtgccgc tgccttcgtt aatatccttt gaataagcca actttgaatc
     5221 acaagacgca taccaaacca aatggtgtcc aagggcgagg agctgttcac cggcgtggtg
     5281 cccatcctgg tggagctgga tggcgacgtg aacggccaca agttcagcgt gcgcggcgag
     5341 ggcgagggcg acgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     5401 ctgcccgtgc cctggcccac cctggtgacc accctgacct acggcgtgca gtgcttcagc
     5461 cgctaccccg atcacatgaa gcagcacgat ttcttcaaga gcgccatgcc cgagggctac
     5521 gtgcaggagc gcaccatcag cttcaaggat gacggcacct acaagacccg cgccgaggtg
     5581 aagttcgagg gcgataccct ggtgaaccgc atcgagctga agggcatcga tttcaaggag
     5641 gatggcaaca tcctgggcca caagctggag tacaacttca acagccacaa cgtgtacatc
     5701 accgccgata agcagaagaa cggcatcaag gccaacttca agatccgcca caatgtggag
     5761 gatggctccg tgcagctggc cgatcactac cagcagaaca cccccatcgg cgacggccca
     5821 gtgctgctgc ccgataacca ctacctgagc acccagagcg tgctgtccaa ggaccccaac
     5881 gagaagcgcg atcacatggt gctgctggag ttcgtgaccg ccgccggcat caccctgggc
     5941 atggatgagc tgtacaagta agcttgatcc agacatgata agatacattg atgagtttgg
     6001 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6061 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     6121 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcgctgtcag gaggaattct
     6181 agtatgtatg taagttaata aaacccattt ttgcggaaag tagataaaaa aaacattttt
     6241 tttttttact gcactggata tcattgaact tatctgatca gttttaaatt tacttcgatc
     6301 caagggtatt tgatgtacca ggttctttcg attacctctc actcaaaatg acattccact
     6361 caaagtcagc gctgtttgcc tccttctctg tccacagaaa tatcgccgtc actttcgccg
     6421 ctgcgtccgc tatctctttc gccaccgttt gtagcgttac gtagcgtcaa tgtccgcctt
     6481 cagttgcatt ttgtcagcgg tttcgtgacg aagctccaag cggtttacgc catcaattaa
     6541 acacaaagtg ctgtgccaaa actcctctcg cttcttattt ttgtttgttt tttgagtgat
     6601 tggggtggtg attggttttg ggtgggtaag caggggaaag tgtgaaaaat cccggcaatg
     6661 ggccaagagg atcaggagct attaattcgc ggaggcagca aacacccatc tgccgagcat
     6721 ctgaacaatg tgagtagtac atgtgcatac atcttaagtt cacttgatct ataggaactg
     6781 cgattgcaac atcaaattgt ctgcggcgtg agaactgcga cccacaaaaa tcccaaaccg
     6841 caattgcaca aacaaatagt gacacgaaac agattattct ggtagctgtt ctcgctatat
     6901 aagacaattt ttgagatcat atcatgatca agacatctaa aggcattcat tttcgactat
     6961 attctttttt acaaaaaata taacaaccag atattttaag ctgatcctag atgcacaaaa
     7021 aataaataaa agtataaacc tacttcgtag gatacttcgg ggtacttttt gttcggggtt
     7081 agatgagcat aacgcttgta gttgatattt gagatcccct atcattgcag ggtgacagcg
     7141 gagcggcttc gcagagctgc attaaccagg gcttcgggca ggccaaaaac tacggcacgc
     7201 tccggccacc cagtccgccg gaggactccg gttcagggag cggccaacta gccgagaacc
     7261 tcacctatgc ctggcacaat atggacatct ttggggcggt caatcagccg ggctccggat
     7321 ggcggcagct ggtcaaccgg acacgcggac tattctgcaa cgagcgacac ataccggcgc
     7381 ccaggaaaca tttgctcaag aacggtgagt ttctattcgc agtcggctga tctgtgtgaa
     7441 atcttaataa agggtccaat taccaatttg aaactcagtt tgcggcgtgg cctatccggg
     7501 cgaacttttg gccgtgatgg gcagttccgg tgccggaaag acgaccctgc tgaatgccct
     7561 tgcctttcga tcgccgcagg gcatccaagt atcgccatcc gggatgcgac tgctcaatgg
     7621 ccaacctgtg gacgccaagg agatgcaggc caggtgcgcc tatgtccagc aggatgacct
     7681 ctttatcggc tccctaacgg ccagggaaca cctgattttc caagccatgg tgcggatgcc
     7741 acgacatctg acctatcggc agcgagtggc ccgcgtggat caggtgatcc aggagctttc
     7801 gctcagcaaa tgtcagcaca cgatcatcgg tgtgcccggc agggtgaaag gtctgtccgg
     7861 cggagaaagg aagcgtctgg cattcgcctc cgaggctcta accgatccgc cgcttctgat
     7921 ctgcgatgag cccacctccg gactggactc ctttaccgcc cacagcgtcg tccaggtgct
     7981 gaagaagctg tcgcagaagg gcaagaccgt catcctgacc attcatcagc cgtcttccga
     8041 gctgtttgag ctctttgaca agatccttct gatggccgag ggcagggtag ctttcttggg
     8101 cactcccagc gaagccgtcg acttcttttc ctagtgagtt cgatgtgttt attaagggta
     8161 tctagtatta cataacatct caactcctat ccagcgtggg tgcccagtgt cctaccaact
     8221 acaatccggc ggacttttac gtacaggtgt tggccgttgt gcccggacgg gagatcgagt
     8281 cccgtgatcg gatcgccaag atatgcgaca attttgccat tagcaaagta gcccgggata
     8341 tggagcagtt gttggccacc aaaaatctgg agaagccact ggagcagccg gagaatgggt
     8401 acacctacaa ggccacctgg ttcatgcagt tccgggcggt cctgtggcga tcctggctgt
     8461 cggtgctcaa ggaaccactc ctcgtaaaag tgcgacttat tcagacaacg gtgagtggtt
     8521 ccagtggaaa caaatgatat aacgcttaca attcttggaa acaaattcgc tagattttag
     8581 atagaattgc ctgattccac acccttctta gtttttttca atgagatgta tagtttatag
     8641 ttttgcagaa gataaataaa tttcatttaa ctcgcgaata ttaatgagat gcgagtaaca
     8701 ttttaatttg cagatggttg ccatcttgat tggcctcatc tttttgggcc aacaactcac
     8761 gcaagtgggt gtgatgaata tcaacggagc catcttcctc ttcctgacca acatgacctt
     8821 tcaaaacgtc tttgccacga taaatgtaag tcatgtttag aatacatttg catttcaata
     8881 atttactaac tttctaatga atcgattcga tttaggtgtt cacctcagag ctgccagttt
     8941 ttatgaggga ggcccgaagt cgactttatc gctgtgacac atactttctg ggcaaaacga
     9001 ttgccgaatt gccgcttttt ctcacagtgc cactggtctt cacggcgatt gcctatccga
     9061 tgatcggact gcgggccgga gtgctgcact tcttcaactg cctggcgctg gtcactctgg
     9121 tggccaatgt gtcaacgtcc ttcggatatc taatatcctg cgccagctcc tcgacctcga
     9181 tggcgctgtc tgtgggtccg ccggttatca taccattcct gctctttggc ggcttcttct
     9241 tgaactcggg ctcggtgcca gtatacctca aatggttgtc gtacctctca tggttccgtt
     9301 acgccaacga gggtctgctg atta
//
