LOCUS       sequence-329667-        9327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Blasticidin resistant 5xUAS 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 9327)
  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 9327)
  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-sfGFP-MiniWhite
FEATURES             Location/Qualifiers
     source          1..9327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..615,686..817,1021..1336,1398..2052,
                     2127..2400,2802..2873))
                     /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     3325..3343
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     polyA_signal    3431..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3468..3487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3522..3541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5152..5173)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    5600..5669
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     5726..5745
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5939..5961
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5953..5970
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5954..5970
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5977..5996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5977..5995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5998..6054
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(6061..6079)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6062..6079)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(6133..6180)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(6204..6659)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6341..6362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6553..6572
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(6714..7505)
                     /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      7973..8561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8462..8481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctactccttg cgtcgggccc gaagtcttag agccagatat gcgagcaccc ggaagctcac
       61 gatgagaatg gccagaccca cgtagtccag cggcagatcg gcggcggaga agttaagtgt
      121 ctccaggatg accttgcccg aactggggca cgtggtgttc gacgatgtgc agctaatttc
      181 gcccggctcc acgtccgccc attggttaat cagcagaccc tcgttggcgt aacggaacca
      241 tgagaggtac gacaaccatt tgaggtatac tggcaccgag cccgagttca agaagaagcc
      301 gccaaagagc aggaatggta tgataaccgg cggacccaca gacagcgcca tcgaggtcga
      361 ggagctggcg caggatatta gatatccgaa ggacgttgac acattggcca ccagagtgac
      421 cagcgccagg cagttgaaga agtgcagcac tccggcccgc agtccgatca tcggataggc
      481 aatcgccgtg aagaccagtg gcactgtgag aaaaagcggc aattcggcaa tcgttttgcc
      541 cagaaagtat gtgtcacagc gataaagtcg acttcgggcc tccctcataa aaactggcag
      601 ctctgaggtg aacacctaaa tcgaatcgat tcattagaaa gttagtaaat tattgaaatg
      661 caaatgtatt ctaaacatga cttacattta tcgtggcaaa gacgttttga aaggtcatgt
      721 tggtcaggaa gaggaagatg gctccgttga tattcatcac acccacttgc gtgagttgtt
      781 ggcccaaaaa gatgaggcca atcaagatgg caaccatctg caaattaaaa tgttactcgc
      841 atctcattaa tattcgcgag ttaaatgaaa tttatttatc ttctgcaaaa ctataaacta
      901 tacatctcat tgaaaaaaac taagaagggt gtggaatcag gcaattctat ctaaaatcta
      961 gcgaatttgt ttccaagaat tgtaagcgtt atatcatttg tttccactgg aaccactcac
     1021 cgttgtctga ataagtcgca cttttacgag gagtggttcc ttgagcaccg acagccagga
     1081 tcgccacagg accgcccgga actgcatgaa ccaggtggcc ttgtaggtgt acccattctc
     1141 cggctgctcc agtggcttct ccagattttt ggtggccaac aactgctcca tatcccgggc
     1201 tactttgcta atggcaaaat tgtcgcatat cttggcgatc cgatcacggg actcgatctc
     1261 ccgtccgggc acaacggcca acacctgtac gtaaaagtcc gccggattgt agttggtagg
     1321 acactgggca cccacgctgg ataggagttg agatgttatg taatactaga tacccttaat
     1381 aaacacatcg aactcactag gaaaagaagt cgacggcttc gctgggagtg cccaagaaag
     1441 ctaccctgcc ctcggccatc agaaggatct tgtcaaagag ctcaaacagc tcggaagacg
     1501 gctgatgaat ggtcaggatg acggtcttgc ccttctgcga cagcttcttc agcacctgga
     1561 cgacgctgtg ggcggtaaag gagtccagtc cggaggtggg ctcatcgcag atcagaagcg
     1621 gcggatcggt tagagcctcg gaggcgaatg ccagacgctt cctttctccg ccggacagac
     1681 ctttcaccct gccgggcaca ccgatgatcg tgtgctgaca tttgctgagc gaaagctcct
     1741 ggatcacctg atccacgcgg gccactcgct gccgataggt cagatgtcgt ggcatccgca
     1801 ccatggcttg gaaaatcagg tgttccctgg ccgttaggga gccgataaag aggtcatcct
     1861 gctggacata ggcgcacctg gcctgcatct ccttggcgtc cacaggttgg ccattgagca
     1921 gtcgcatccc ggatggcgat acttggatgc cctgcggcga tcgaaaggca agggcattca
     1981 gcagggtcgt ctttccggca ccggaactgc ccatcacggc caaaagttcg cccggatagg
     2041 ccacgccgca aactgagttt caaattggta attggaccct ttattaagat ttcacacaga
     2101 tcagccgact gcgaatagaa actcaccgtt cttgagcaaa tgtttcctgg gcgccggtat
     2161 gtgtcgctcg ttgcagaata gtccgcgtgt ccggttgacc agctgccgcc atccggagcc
     2221 cggctgattg accgccccaa agatgtccat attgtgccag gcataggtga ggttctcggc
     2281 tagttggccg ctccctgaac cggagtcctc cggcggactg ggtggccgga gcgtgccgta
     2341 gtttttggcc tgcccgaagc cctggttaat gcagctctgc gaagccgctc cgctgtcacc
     2401 ctgcaatgat aggggatctc aaatatcaac tacaagcgtt atgctcatct aaccccgaac
     2461 aaaaagtacc ccgaagtatc ctacgaagta ggtttatact tttatttatt ttttgtgcat
     2521 ctaggatcag cttaaaatat ctggttgtta tattttttgt aaaaaagaat atagtcgaaa
     2581 atgaatgcct ttagatgtct tgatcatgat atgatctcaa aaattgtctt atatagcgag
     2641 aacagctacc agaataatct gtttcgtgtc actatttgtt tgtgcaattg cggtttggga
     2701 tttttgtggg tcgcagttct cacgccgcag acaatttgat gttgcaatcg cagttcctat
     2761 agatcaagtg aacttaagat gtatgcacat gtactactca cattgttcag atgctcggca
     2821 gatgggtgtt tgctgcctcc gcgaattaat agctcctgat cctcttggcc cattgccggg
     2881 atttttcaca ctttcccctg cttacccacc caaaaccaat caccacccca atcactcaaa
     2941 aaacaaacaa aaataagaag cgagaggagt tttggcacag cactttgtgt ttaattgatg
     3001 gcgtaaaccg cttggagctt cgtcacgaaa ccgctgacaa aatgcaactg aaggcggaca
     3061 ttgacgctac gtaacgctac aaacggtggc gaaagagata gcggacgcag cggcgaaagt
     3121 gacggcgata tttctgtgga cagagaagga ggcaaacagc gctgactttg agtggaatgt
     3181 cattttgagt gagaggtaat cgaaagaacc tggtacatca aatacccttg gatcgaagta
     3241 aatttaaaac tgatcagata agttcaatga tatccagtgc agtaaaaaaa aaaaatgttt
     3301 tttttatcta ctttccgcaa aaatgggttt tattaactta catacatact agaattcctc
     3361 ctgacagcga aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3421 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3541 caatgtatct tatcatgtct ggatcaagct tacttgtaca gctcatccat gcccagggtg
     3601 atgccggcgg cggtcacgaa ctccagcagc accatgtgat cgcgcttctc gttggggtcc
     3661 ttggacagca cgctctgggt gctcaggtag tggttatcgg gcagcagcac tgggccgtcg
     3721 ccgatggggg tgttctgctg gtagtgatcg gccagctgca cggagccatc ctccacattg
     3781 tggcggatct tgaagttggc cttgatgccg ttcttctgct tatcggcggt gatgtacacg
     3841 ttgtggctgt tgaagttgta ctccagcttg tggcccagga tgttgccatc ctccttgaaa
     3901 tcgatgccct tcagctcgat gcggttcacc agggtatcgc cctcgaactt cacctcggcg
     3961 cgggtcttgt aggtgccgtc atccttgaag ctgatggtgc gctcctgcac gtagccctcg
     4021 ggcatggcgc tcttgaagaa atcgtgctgc ttcatgtgat cggggtagcg gctgaagcac
     4081 tgcacgccgt aggtcagggt ggtcaccagg gtgggccagg gcacgggcag cttgccggtg
     4141 gtgcagatga acttcagggt cagcttgccg ttggtggcgt cgccctcgcc ctcgccgcgc
     4201 acgctgaact tgtggccgtt cacgtcgcca tccagctcca ccaggatggg caccacgccg
     4261 gtgaacagct cctcgccctt ggacaccatt tggtttggta tgcgtcttgt gattcaaagt
     4321 tggcttattc aaaggatatt aacgaaggca gcggcacgtc tgctcggctc gaacattcat
     4381 tcacaactga tgctctcagc caccccgcgc ccttttatac cgctgcgctc gatccccggg
     4441 cgagctcggg acggaggaca gtactccgct cggaggacag tactccgctc ggaggacagt
     4501 actccgctcg gaggacagta ctccgctcgg aggacagtac tccgctcctg acagcggaac
     4561 aaacgaccca acacccgtgc gttttattct gtctttttat tgccaagctt agttacgggt
     4621 atacttcagg ggaatcagtt cctcaatggt ggtcttgacc agcttgccgt tcatctcaat
     4681 cagcacgaag caatcggggg cgtaatcgct aatcagctca cggcacattc cacacgggga
     4741 cacgacacgg atgctacggt ccacctcatc ggagtatggg tgacgcacgg ccacaatggt
     4801 atcaaaatcc ttctggccgt tgctcacggc actgccaatg gcaatggcct cggcacacac
     4861 ggtcacgcgt ccgatgtagg cctcaatgtg cacggcactg ataatctcgc ccgtcttggt
     4921 gcgaatggca gctcccacgt ggtgcttgtt atcctcgtag agcatcgtaa tcttctcggt
     4981 agccacctcc accagttcca ggtcctgctg gctgatgttg aaggttttca ttttgttttc
     5041 ctccttatgt taagcttact cagttattat atcataaata tctgtgtcaa gaataaactc
     5101 ccacatggat tcgagtattg agtgttctct tcttgtattc aataattact tcttggcaga
     5161 tttcagtagt tgcagttgat ttacttggtt gctggttact tttaattgat tcactttaac
     5221 ttgcacttta ctgcagattg tttagcttgt tcagctgcgc ttgtttattt gcttagcttt
     5281 cgcttagcga cgtgttcact ttgcttgttt gaattgaatt gtcgctccgt agacgaagcg
     5341 cctctattta tactccggcg ctcttttcgc gaacattcga ggcgcgctct ctcgaaccaa
     5401 cgagagcagt atgccgttta ctgtgtgaca gagtgagaga gcattagtgc agagagggag
     5461 cgacccaaaa agaaaagaga gaataacgaa taacggccag agaaatttct cgagttttct
     5521 ttctgccaaa caaatgacct accacaataa ccagtttgtt ttgggattct agctcctgac
     5581 agcgcatcat gatggaccag atgggtgagg tggagtacgc gcccggggag cccaagggca
     5641 cgccctggca cccgcaccgc ggcttcgaga cagtgaccta catcgtcgac caattcctcc
     5701 ctgggatccg gagagagacc ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5761 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgtcattcg
     5821 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5881 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     5941 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     6001 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttgagtat
     6061 tctatagtgt cacctaaata gcttggcgta atcatggtca tgatctgcct cctgggttta
     6121 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     6181 aggggtctct cgctgatccg ttaaaattgt aagcgttaat attttgttaa aattcgcgtt
     6241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6361 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgctgg
     6421 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6481 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6541 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6601 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     6661 acatgaagtt ttaaatcaat ctaaagtata tatgtgtaac attggtctag tgattatttg
     6721 ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg
     6781 cgcgaggcca aacgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg
     6841 ggctgatact gggccggcag gcgctccatt gcccagtcgg cagctacatc cttcggcgcg
     6901 attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca
     6961 tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat
     7021 agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg
     7081 ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg
     7141 aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct
     7201 ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga
     7261 atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc
     7321 gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc
     7381 aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg
     7441 cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc
     7501 ctcataatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc
     7561 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
     7621 gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca
     7681 ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac
     7741 agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt
     7801 gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg catttctgtc ctggctggtc
     7861 acaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7921 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8101 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     8341 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     8401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8641 ctgattctgt ggataaccgt gtcaagcgat gcatctacac aaggaacaaa cactggatgt
     8701 cactttcagt tcaaattgta acgctaatca ctccgaacag gtcacaaaaa attaccttaa
     8761 aaagtcataa tattaaatta gaataaatat agctgtgagg gaaatatata caaatatatt
     8821 ggagcaaata aattgtacat acaaatattt attactaatt tctattgagc cgaaatgaac
     8881 cactcggaac catttgagcg aaccgaatcg cgcggaacta acgacagtcg ctccaaggtc
     8941 gtcgaacaaa aggtgaatgt gttgcggaga gcgggtggga gacagcgaaa gagcaactac
     9001 gaaacgtggt gtggtggagg tgaattatga agagggcgcg cgatttgaaa agtatgtata
     9061 taaaaaatat atcccggtgt tttatgtagc gataaacgag tttttgatgt aaggtatgca
     9121 ggtgtgtaag tcttttggtt agaagacaaa tccaaagtct acttgtgggg atgttcgaag
     9181 gggaaatact tgtattctat aggtcatatc ttgtttttat tggcacaaat ataattacat
     9241 tagctttttg agggggcaat aaacagtaaa cacgatggta ataatggtaa aaaaaaaaaa
     9301 caagcagtta tttcggatat atgtcgg
//
