LOCUS       sequence-328534-        8033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xQUAS EBFP2 GMR-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 8033)
  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 8033)
  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-GMRWhite
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2466..2600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2873..3461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2953..2972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3929..4720
                     /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      4775..5230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4862..4881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5072..5093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5254..5301
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5355..5373
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5355..5372
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    5380..5436
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5438..5457)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5439..5457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5464..5481)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5464..5480)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5473..5495)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5689..5708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    5765..5834
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     6261..6282
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     polyA_signal    7866..8000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gttcaggggg aggtgtggga ggttttttcg ctgtcaggag cgtacgacta gcacctgact
       61 gtcgagccca gtggaaaccc ttgaaatgcc tttaagtcga gcccagtgga aacccttgaa
      121 atgcctttaa gtcgagccca gtggaaaccc ttgaaatgcc tttaagtcga gcccagtgga
      181 aacccttgaa atgcctttaa gtcgagtagc agtcgaggta ggtacgtcat gcccctcccg
      241 agctcgcccg gggatcgagc gcagcggtat aaaagggcgc ggggtggctg agagcatcag
      301 ttgtgaatga atgttcgagc cgagcagacg tgccgctgcc ttcgttaata tcctttgaat
      361 aagccaactt tgaatcacaa gacgcatacc aaaccaaatg ggacaggagg accaggagct
      421 gctgatccgc ggaggctcca agcacccaag cgccgagcac ctgaacaacg gcgatagcgg
      481 agccgccagc cagtcgtgca tcaatcaggg atttggccag gccaagaatt acggaaccct
      541 gctgcccccc agccccccag aggatagcgg aagcggatcc ggacagctgg ccgagaatct
      601 gacgtacgcc tggcataaca tggatatctt cggcgccgtg aaccagccag gcagtggctg
      661 gcgtcagctg gtgaaccgca cccgcggact gttctgcaat gagcgccata tcccagcccc
      721 acgcaagcac ctgctgaaga atgtgtgcgg agtggcctac cccggcgagc tgctggccgt
      781 gatgggaagt agtggcgccg gaaagaccac gctgctgaac gccctggcct tccgcagccc
      841 ccagggcatt caggtgtccc caagcggcat gcgcctgctg aacggccagc ccgtggatgc
      901 caaggagatg caggcccgct gcgcctacgt gcagcaggac gatctgttca tcggcagcct
      961 gacagcccgc gagcatctga tctttcaggc catggtccgc atgccccgcc acctgacata
     1021 ccgtcagcgc gtggcccgcg tggaccaggt catccaggag ctgagcctga gcaagtgcca
     1081 gcacaccatt attggcgtgc caggccgcgt gaagggcctg agtggcggag agcgcaagcg
     1141 cctggccttt gccagcgagg ccctgaccga tccccccctg ctgatttgcg acgagccgac
     1201 cagcggcctg gatagcttca ccgcccactc cgtggtgcag gtgctgaaga agctgagcca
     1261 gaagggaaag acggtcattc tgacgatcca ccagcccagc tcggagctgt tcgagctgtt
     1321 tgataagatt ctgctgatgg ccgagggacg cgtggccttc ctgggaaccc caagcgaggc
     1381 cgtggatttc ttcagctatg tgggcgccca gtgccccacg aattacaacc ccgccgactt
     1441 ctatgtgcaa gtgctggccg tggtgccagg acgcgagatt gagagccgcg atcgcattgc
     1501 caagatctgc gataacttcg ccatctccaa ggtggcccgc gacatggagc agctgctggc
     1561 cacgaagaac ctggagaagc ccctggagca gcccgagaac ggctatacgt ataaggctac
     1621 ctggtttatg cagtttcgcg ccgtgctgtg gcgcagctgg ctgagtgtgc tgaaggagcc
     1681 cctgctggtc aaggtccgcc tgatccagac cacgatggtg gccattctga tcggcctgat
     1741 cttcctggga cagcagctga cgcaggtcgg agtgatgaac atcaatggag ccatttttct
     1801 gtttctgacg aacatgacgt tccagaacgt gttcgccacc atcaacgtgt tcaccagcga
     1861 gctgcccgtg ttcatgcgcg aggcccgcag ccgcctgtac cgctgcgata cctacttcct
     1921 gggaaagacc attgccgagc tgccgctgtt cctgaccgtg cccctggtgt ttaccgccat
     1981 tgcctacccc atgattggcc tgcgtgccgg agtgctgcat ttctttaatt gcctggccct
     2041 ggtgaccctg gtggccaacg tgtcgaccag cttcggctac ctgatcagct gcgcctccag
     2101 cagcaccagc atggccctga gcgtgggacc ccccgtgatt atccccttcc tgctgttcgg
     2161 cggattcttt ctgaactccg gcagcgtgcc cgtgtacctg aagtggctga gctacctgag
     2221 ctggtttcgc tacgccaatg agggcctgct gatcaatcag tgggccgatg tggagcccgg
     2281 cgagatctcc tgcaccagca gcaacacgac ctgccccagc agcggaaagg tcatcctgga
     2341 gacactgaat ttcagtgccg ccgacctgcc actggattat gtgggcctgg ccatcctgat
     2401 tgtgtccttc cgcgtgctgg cctatctggc tctgcgcctg cgcgctcgtc gcaaggagtg
     2461 agcttgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2581 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2641 gggaggtgtg ggaggttttt tcagtatggc aagcgctcga cgaattaaca agcttcgctc
     2701 tatctcgctc tctatctctc tcgatcgctc gtgctaagct taagaccgtg gagaacaagg
     2761 agaaccgctt gacacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2821 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3301 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3541 tatcaaaaag gatcttcacc tagatccttt tgtgaccagc caggacagaa atgcctcgac
     3601 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     3661 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     3721 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     3781 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     3841 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     3901 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
     3961 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     4021 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     4081 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     4141 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     4201 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     4261 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     4321 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     4381 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     4441 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     4501 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtag ctgccgactg
     4561 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     4621 tcttggacaa gaagaagatc gtttggcctc gcgcgcagat cagttggaag aatttgtcca
     4681 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa tcactagacc aatgttacac
     4741 atatatactt tagattgatt taaaacttca tgttacgcgc cctgtagcgg cgcattaagc
     4801 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4861 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4921 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4981 aaacttgatt agggtgatgg ttcacgtagt gggccagcgc cctgatagac ggtttttcgc
     5041 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5101 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5161 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5221 cttacaattt taacggatca gcgagagacc cctgttgaca attaatcatc ggcatagtat
     5281 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgaccat
     5341 gattacgcca agctatttag gtgacactat agaatactca agcttgcatg cctgcaggtc
     5401 gactctagag gatccccggg taccgagctc gaattcgccc tatagtgagt cgtattacaa
     5461 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5521 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5581 tcgcccttcc caacagttgc gcagcctgaa tggcgaatga cgcctgatgc ggtattttct
     5641 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     5701 tgatgccgca tagggtctct ctccggatcc cagggaggaa ttggtcgacg atgtaggtca
     5761 ctgtctcgaa gccgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
     5821 ccacctcacc catctggtcc atcatgatgc gctgtcagga gctagaatcc caaaacaaac
     5881 tggttattgt ggtaggtcat ttgtttggca gaaagaaaac tcgagaaatt tctctggccg
     5941 ttattcgtta ttctctcttt tctttttggg tcgctccctc tctgcactaa tgctctctca
     6001 ctctgtcaca cagtaaacgg catactgctc tcgttggttc gagagagcgc gcctcgaatg
     6061 ttcgcgaaaa gagcgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa
     6121 ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc
     6181 tgaacaagct aaacaatctg cagtaaagtg caagttaaag tgaatcaatt aaaagtaacc
     6241 agcaaccaag taaatcaact gcaactactg aaatctgcca agaagtaatt attgaataca
     6301 agaagagaac actcaatact cgaatccatg tgggagttta ttcttgacac agatatttat
     6361 gatataataa ctgagtaagc ttaacataag gaggaaaaca aaatgaaaac cttcaacatc
     6421 agccagcagg acctggaact ggtggaggtg gctaccgaga agattacgat gctctacgag
     6481 gataacaagc accacgtggg agctgccatt cgcaccaaga cgggcgagat tatcagtgcc
     6541 gtgcacattg aggcctacat cggacgcgtg accgtgtgtg ccgaggccat tgccattggc
     6601 agtgccgtga gcaacggcca gaaggatttt gataccattg tggccgtgcg tcacccatac
     6661 tccgatgagg tggaccgtag catccgtgtc gtgtccccgt gtggaatgtg ccgtgagctg
     6721 attagcgatt acgcccccga ttgcttcgtg ctgattgaga tgaacggcaa gctggtcaag
     6781 accaccattg aggaactgat tcccctgaag tatacccgta actaagcttg gcaataaaaa
     6841 gacagaataa aacgcacggg tgttgggtcg tttgttccgc tgtcaggagg ggtaatcgct
     6901 tatcctcgga taaacaatta tcctcacggg taatcgctta tccgggtaat cgcttatcct
     6961 cgggtaatcg cttatccttt cccgagctcg cccggggatc gagcgcagcg gtataaaagg
     7021 gcgcggggtg gctgagagca tcagttgtga atgaatgttc gagccgagca gacgtgccgc
     7081 tgccttcgtt aatatccttt gaataagcca actttgaatc acaagacgca taccaaacca
     7141 aatggtgtcc aagggcgagg agctgttcac cggcgtggtg cccatcctgg tggagctgga
     7201 tggcgacgtg aacggccaca agttcagcgt gcgcggcgag ggcgagggcg acgccaccaa
     7261 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     7321 cctggtgacc accctgagcc acggcgtgca gtgcttcgcc cgctaccccg atcacatgaa
     7381 gcagcacgat ttcttcaaga gcgccatgcc cgagggctac gtgcaggagc gcaccatttt
     7441 cttcaatgac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgataccct
     7501 ggtgaaccgc atcgagctga agggcgtgga tttcaaggag gatggcaaca tcctgggcca
     7561 caagctggag tacaacttca acagccacaa catctacatc atggccgtga agcagaagaa
     7621 cggcatcaag gtcaacttca agatccgcca caacgtggag gatggctcag tgcagctggc
     7681 cgatcactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgatagcca
     7741 ctacctgagc acccagagcg tgctgtccaa ggatcccaac gagaagcgcg atcacatggt
     7801 gctgctggag ttccgcaccg ccgccggcat caccctgggc atggatgagc tgtacaagta
     7861 agcttgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     7921 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     7981 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt cag
//
