LOCUS       sequence-326306-        9169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant LexA driver vector with Mini-white CDS eye 
            marker. Contains an enhancer grammar GB20 entry point for custom 
            enhancers. Standard cut-and-paste cloning can also be used. Vector 
            uses Purple-White bacteria colony screening for identification of 
            correct cloning .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9169)
  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 9169)
  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-MCS-LexA-VP16-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2473..2607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2880..3468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2960..2979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3936..4727
                     /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      4782..5237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4869..4888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5079..5100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     protein_bind    5360..5429
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        5457..5908
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     5856..5877
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     promoter        6857..6875
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6857..6874
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6886..6903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      7711..7805
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    7812..7868
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(7930..7948)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(8111..8362,8363..8716)
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     CDS             8768..9001
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     polyA_signal    9009..9143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gtttcaggtt cagggggagg tgtgggaggt tttttcgctg tcaggagcgt acgactagca
       61 cctgactgtc gagcccagtg gaaacccttg aaatgccttt aagtcgagcc cagtggaaac
      121 ccttgaaatg cctttaagtc gagcccagtg gaaacccttg aaatgccttt aagtcgagcc
      181 cagtggaaac ccttgaaatg cctttaagtc gagtagcagt cgaggtaggt acgtcatgcc
      241 cctcccgagc tcgcccgggg atcgagcgca gcggtataaa agggcgcggg gtggctgaga
      301 gcatcagttg tgaatgaatg ttcgagccga gcagacgtgc cgctgccttc gttaatatcc
      361 tttgaataag ccaactttga atcacaagac gcataccaaa ccaaatggga caggaggacc
      421 aggagctgct gatccgcgga ggctccaagc acccaagcgc cgagcacctg aacaacggcg
      481 atagcggagc cgccagccag tcgtgcatca atcagggatt tggccaggcc aagaattacg
      541 gaaccctgct gccccccagc cccccagagg atagcggaag cggatccgga cagctggccg
      601 agaatctgac gtacgcctgg cataacatgg atatcttcgg cgccgtgaac cagccaggca
      661 gtggctggcg tcagctggtg aaccgcaccc gcggactgtt ctgcaatgag cgccatatcc
      721 cagccccacg caagcacctg ctgaagaatg tgtgcggagt ggcctacccc ggcgagctgc
      781 tggccgtgat gggaagtagt ggcgccggaa agaccacgct gctgaacgcc ctggccttcc
      841 gcagccccca gggcattcag gtgtccccaa gcggcatgcg cctgctgaac ggccagcccg
      901 tggatgccaa ggagatgcag gcccgctgcg cctacgtgca gcaggacgat ctgttcatcg
      961 gcagcctgac agcccgcgag catctgatct ttcaggccat ggtccgcatg ccccgccacc
     1021 tgacataccg tcagcgcgtg gcccgcgtgg accaggtcat ccaggagctg agcctgagca
     1081 agtgccagca caccattatt ggcgtgccag gccgcgtgaa gggcctgagt ggcggagagc
     1141 gcaagcgcct ggcctttgcc agcgaggccc tgaccgatcc ccccctgctg atttgcgacg
     1201 agccgaccag cggcctggat agcttcaccg cccactccgt ggtgcaggtg ctgaagaagc
     1261 tgagccagaa gggaaagacg gtcattctga cgatccacca gcccagctcg gagctgttcg
     1321 agctgtttga taagattctg ctgatggccg agggacgcgt ggccttcctg ggaaccccaa
     1381 gcgaggccgt ggatttcttc agctatgtgg gcgcccagtg ccccacgaat tacaaccccg
     1441 ccgacttcta tgtgcaagtg ctggccgtgg tgccaggacg cgagattgag agccgcgatc
     1501 gcattgccaa gatctgcgat aacttcgcca tctccaaggt ggcccgcgac atggagcagc
     1561 tgctggccac gaagaacctg gagaagcccc tggagcagcc cgagaacggc tatacgtata
     1621 aggctacctg gtttatgcag tttcgcgccg tgctgtggcg cagctggctg agtgtgctga
     1681 aggagcccct gctggtcaag gtccgcctga tccagaccac gatggtggcc attctgatcg
     1741 gcctgatctt cctgggacag cagctgacgc aggtcggagt gatgaacatc aatggagcca
     1801 tttttctgtt tctgacgaac atgacgttcc agaacgtgtt cgccaccatc aacgtgttca
     1861 ccagcgagct gcccgtgttc atgcgcgagg cccgcagccg cctgtaccgc tgcgatacct
     1921 acttcctggg aaagaccatt gccgagctgc cgctgttcct gaccgtgccc ctggtgttta
     1981 ccgccattgc ctaccccatg attggcctgc gtgccggagt gctgcatttc tttaattgcc
     2041 tggccctggt gaccctggtg gccaacgtgt cgaccagctt cggctacctg atcagctgcg
     2101 cctccagcag caccagcatg gccctgagcg tgggaccccc cgtgattatc cccttcctgc
     2161 tgttcggcgg attctttctg aactccggca gcgtgcccgt gtacctgaag tggctgagct
     2221 acctgagctg gtttcgctac gccaatgagg gcctgctgat caatcagtgg gccgatgtgg
     2281 agcccggcga gatctcctgc accagcagca acacgacctg ccccagcagc ggaaaggtca
     2341 tcctggagac actgaatttc agtgccgccg acctgccact ggattatgtg ggcctggcca
     2401 tcctgattgt gtccttccgc gtgctggcct atctggctct gcgcctgcgc gctcgtcgca
     2461 aggagtgagc ttgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     2521 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2581 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2641 gttcaggggg aggtgtggga ggttttttca gtatggcaag cgctcgacga attaacaagc
     2701 ttcgctctat ctcgctctct atctctctcg atcgctcgtg ctaagcttaa gaccgtggag
     2761 aacaaggaga accgcttgac acggttatcc acagaatcag gggataacgc aggaaagaac
     2821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3301 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3541 atgagattat caaaaaggat cttcacctag atccttttgt gaccagccag gacagaaatg
     3601 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     3661 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
     3721 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     3781 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     3841 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     3901 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
     3961 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     4021 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     4081 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     4141 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     4201 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     4261 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     4321 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     4381 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     4441 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     4501 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtagctg
     4561 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     4621 aggcttatct tggacaagaa gaagatcgtt tggcctcgcg cgcagatcag ttggaagaat
     4681 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatca ctagaccaat
     4741 gttacacata tatactttag attgatttaa aacttcatgt tacgcgccct gtagcggcgc
     4801 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4861 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4921 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4981 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt
     5041 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5101 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5161 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5221 attaacgctt acaattttaa cggatcagcg cagctgcccg ggtctagagt cgacgcatgc
     5281 ctcctgacag cgagatgtcc taatatggac atcctctccg gatcccaggg aggaattggt
     5341 cgacgatgta ggtcactgtc tcgaagccgc ggtgcgggtg ccagggcgtg cccttgggct
     5401 ccccgggcgc gtactccacc tcacccatct ggtccatcat gatgcgctgt caggagctag
     5461 aatcccaaaa caaactggtt attgtggtag gtcatttgtt tggcagaaag aaaactcgag
     5521 aaatttctct ggccgttatt cgttattctc tcttttcttt ttgggtcgct ccctctctgc
     5581 actaatgctc tctcactctg tcacacagta aacggcatac tgctctcgtt ggttcgagag
     5641 agcgcgcctc gaatgttcgc gaaaagagcg ccggagtata aatagaggcg cttcgtctac
     5701 ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc gctaagcgaa agctaagcaa
     5761 ataaacaagc gcagctgaac aagctaaaca atctgcagta aagtgcaagt taaagtgaat
     5821 caattaaaag taaccagcaa ccaagtaaat caactgcaac tactgaaatc tgccaagaag
     5881 taattattga atacaagaag agaactctca atactcgaat ccatgtggga gtttattctt
     5941 gacacagata tttatgatat aataactgag taagcttaac ataaggagga aaacaaaatg
     6001 attgagcagg atggactgca cgctggctcg ccagctgcct gggtggagcg tctgttcggc
     6061 tacgattggg cccagcagac catcggctgc tccgatgctg ccgtgttccg tctgtcggcc
     6121 cagggacgcc ccgtgctgtt cgtcaagacc gacctgagtg gagccctgaa cgagctccag
     6181 gatgaggcag ctcgtctgag ctggctggcc acaacaggag tgccgtgtgc tgccgtgctg
     6241 gatgtggtga ccgaagctgg acgcgattgg ctgctgctgg gcgaggtgcc aggccaggac
     6301 ctgctgtcca gccatctggc cccagccgag aaggtgtcca ttatggccga tgctatgcgt
     6361 cgcctgcaca ccctggaccc agccacctgc ccctttgacc accaggccaa gcaccgtatt
     6421 gagcgtgccc gaacccgtat ggaggctgga ctggtggacc aggatgacct ggatgaggag
     6481 caccagggcc tggctccagc cgagctgttt gcccgtctga aggcccgtat gcccgatggc
     6541 gaggacctgg tggtgaccca cggcgacgcc tgcctgccca acattatggt ggagaacgga
     6601 cgcttcagtg gcttcattga ttgtggacgc ctgggcgtgg ccgaccgtta ccaggatatc
     6661 gccctggcca cccgagatat cgccgaggag ctgggaggcg agtgggccga ccgattcctg
     6721 gtgctctacg gcattgcagc cccagatagc cagcgtattg ccttctaccg tctgctggac
     6781 gagttcttct aagcttggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     6841 gttccgctgt caggagattt aggtgacact atagacttcc gcttcctcgc tcactgactc
     6901 gctgcgctcg gtcgttcggc tgtcgggaga gagaccaatc catgtgggag tttattcttg
     6961 acacagatat ttatgatata ataactgagt aagcttaaca taaggaggaa aacaaaatgg
     7021 cgagcttggt taagaaagat atgtgtgtta agatgacgat ggaaggtact gtgaacggtt
     7081 atcactttaa gtgcgttggc gagggtgaag gcaagccgtt cgagggcacg cagaacatgc
     7141 gcattcgtgt caccgagggc ggtccgctgc cttttgcatt cgacatcctg gccccgtgct
     7201 gtatgtacgg ctctaagacc ttcattaaac acgtgagcgg tatcccggat tactttaaag
     7261 agtcctttcc agagggcttc acttgggaac gtacccagat ttttgaggac ggtggtgttc
     7321 tgaccgcgca ccaagacacc agcctggaag gtaattgcct gatctataaa gtgaaggttc
     7381 tgggtaccaa tttcccggcg aatggtccgg tgatgcaaaa gaaaaccgcg ggttgggagc
     7441 cgtgcgtcga gatgctgtat ccgcgtgacg gcgtcttgtg tggtcagagc ttgatggcgc
     7501 tgaagtgcac cgatggcaat catctgacca gccacctgcg cacgacgtat cgtagccgta
     7561 aaccgagcaa cgccgttaac atgccggagt tccattttgg tgaccatcgc atcgaaatcc
     7621 tgaaagctga gcagggcaaa ttctacgaac aatacgaatc ggctgtcgca cgttacagcg
     7681 atgtgccgga aaaagcgacg taatgagctt gactcctgtt gatagatcca gtaatgacct
     7741 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     7801 agaattactt caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
     7861 tcgaattcgg tctcttcccg cttgtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     7921 cgcatcaggc ctatagtgag tcgtattatc ccgagctcgc ccggggatcg agcgcagcgg
     7981 tataaaaggg cgcggggtgg ctgagagcat cagttgtgaa tgaatgttcg agccgagcag
     8041 acgtgccgct gccttcgtta atatcctttg aataagccaa ctttgaatca caagacgcat
     8101 accaaaccaa atgaaggctc tcacggcccg acaacaggaa gtttttgatt tgatacggga
     8161 tcatatatcc caaacgggta tgcctccgac ccgcgcagag atagcacagc gactgggctt
     8221 tcgatcgcct aacgccgcgg aggagcactt gaaggcactg gcccgcaagg gtgtcattga
     8281 aatcgtgtcc ggtgcgagcc gcggaatccg gctgttgcag gaggaggaag agggcctgcc
     8341 actggtggga cgcgtggccg ctggcgagcc gctgctggcc cagcaacaca tagagggaca
     8401 ctatcaggtg gacccctcct tgtttaagcc aaatgctgat ttcctgttgc gggtgtcggg
     8461 aatgtccatg aaggacatcg gtattatgga tggtgacctc ctcgccgtcc ataagacaca
     8521 ggatgtcagg aacggccagg tagtcgttgc caggatagac gatgaggtca ctgtgaaacg
     8581 cctcaagaag caaggcaata aggtcgagct gctgccggag aatagcgagt tcaagccgat
     8641 cgtggtggat ctgcgacagc agtcctttac tatcgagggc ttggccgtgg gtgtgatccg
     8701 caacggagat tggctgagtg gcggaggttc cggtggcggc agcggcgggg gttcgggagg
     8761 tggctcggcc cccccgaccg atgtcagcct gggggacgag ctccacttag acggcgagga
     8821 cgtggccatg gcgcatgccg acgcgctaga cgatttcgat ctggacatgt tgggggacgg
     8881 ggattccccg gggccgggat ttacccccca cgactccgcc ccctacggcg ctctggatac
     8941 ggccgacttc gagtttgagc agatgtttac cgatgccctt ggaattgacg agtacggtgg
     9001 gtaagcttga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     9061 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     9121 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttat
//
