LOCUS       sequence-328535-        7103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin resistance vector with attB and eye marker. Contains an 
            Alpha GB cloning entry point for addition of custom assembly uses 
            blue-white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7103)
  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 7103)
  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: pPuroR-GMRWhite
FEATURES             Location/Qualifiers
     source          1..7103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(59..78)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(60..78)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(85..102)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(85..101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(94..116)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(310..329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    386..455
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        483..934
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     882..903
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     polyA_signal    4148..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4172..4191)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4226..4245
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4597..5185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4677..4696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5653..6444
                     /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      6499..6954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6586..6605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6796..6817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6978..7025
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7079..7097
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7079..7096
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcgcc
       61 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      121 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      181 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      241 acgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
      301 cactctcagt acaatctgct ctgatgccgc atagggtctc tctccggatc ccagggagga
      361 attggtcgac gatgtaggtc actgtctcga agccgcggtg cgggtgccag ggcgtgccct
      421 tgggctcccc gggcgcgtac tccacctcac ccatctggtc catcatgatg cgctgtcagg
      481 agctagaatc ccaaaacaaa ctggttattg tggtaggtca tttgtttggc agaaagaaaa
      541 ctcgagaaat ttctctggcc gttattcgtt attctctctt ttctttttgg gtcgctccct
      601 ctctgcacta atgctctctc actctgtcac acagtaaacg gcatactgct ctcgttggtt
      661 cgagagagcg cgcctcgaat gttcgcgaaa agagcgccgg agtataaata gaggcgcttc
      721 gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct
      781 aagcaaataa acaagcgcag ctgaacaagc taaacaatct gcagtaaagt gcaagttaaa
      841 gtgaatcaat taaaagtaac cagcaaccaa gtaaatcaac tgcaactact gaaatctgcc
      901 aagaagtaat tattgaatac aagaagagaa ctctcaatac tcgaatccat gtgggagttt
      961 attcttgaca cagatattta tgatataata actgagtaag cttaacataa ggaggaaaac
     1021 aaaatgaccg agtataagcc cacggtgcgt ctggccaccc gtgatgatgt gccccgtgcc
     1081 gtgcgaaccc tggctgctgc cttcgccgat tacccagcca cccgtcacac cgtggaccca
     1141 gaccgtcaca ttgagcgtgt gaccgagctc caggagctgt tcctgacacg cgtgggcctg
     1201 gatatcggca aggtctgggt ggccgatgat ggagctgccg tggccgtgtg gaccaccccc
     1261 gagagcgtgg aggctggagc cgtgttcgcc gagattggac cccgtatggc cgagctgagt
     1321 ggctcccgtc tggctgccca gcagcagatg gagggcctgc tggccccaca ccgtccaaag
     1381 gagcccgtct ggttcctggc caccgtgggc gtgagccccg accaccaggg caagggcctg
     1441 ggcagtgccg tggtgctgcc aggagtggaa gctgccgagc gtgctggagt gccagccttc
     1501 ctggaaacca gtgccccacg caatctgccc ttctacgagc gtctgggctt caccgtgacg
     1561 gccgatgtgg aggtgcccga gggcccacgc acctggtgta tgacccgtaa gccaggagcc
     1621 taagcttggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttccgctg
     1681 tcaggagcgt acgactagca cctgactgtc gagcccagtg gaaacccttg aaatgccttt
     1741 aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg gaaacccttg
     1801 aaatgccttt aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg
     1861 gaaacccttg aaatgccttt aagtcgagta gcagtcgagg taggtacgtc atgcccctcc
     1921 cgagctcgcc cggggatcga gcgcagcggt ataaaagggc gcggggtggc tgagagcatc
     1981 agttgtgaat gaatgttcga gccgagcaga cgtgccgctg ccttcgttaa tatcctttga
     2041 ataagccaac tttgaatcac aagacgcata ccaaaccaaa tgggacagga ggaccaggag
     2101 ctgctgatcc gcggaggctc caagcaccca agcgccgagc acctgaacaa cggcgatagc
     2161 ggagccgcca gccagtcgtg catcaatcag ggatttggcc aggccaagaa ttacggaacc
     2221 ctgctgcccc ccagcccccc agaggatagc ggaagcggat ccggacagct ggccgagaat
     2281 ctgacgtacg cctggcataa catggatatc ttcggcgccg tgaaccagcc aggcagtggc
     2341 tggcgtcagc tggtgaaccg cacccgcgga ctgttctgca atgagcgcca tatcccagcc
     2401 ccacgcaagc acctgctgaa gaatgtgtgc ggagtggcct accccggcga gctgctggcc
     2461 gtgatgggaa gtagtggcgc cggaaagacc acgctgctga acgccctggc cttccgcagc
     2521 ccccagggca ttcaggtgtc cccaagcggc atgcgcctgc tgaacggcca gcccgtggat
     2581 gccaaggaga tgcaggcccg ctgcgcctac gtgcagcagg acgatctgtt catcggcagc
     2641 ctgacagccc gcgagcatct gatctttcag gccatggtcc gcatgccccg ccacctgaca
     2701 taccgtcagc gcgtggcccg cgtggaccag gtcatccagg agctgagcct gagcaagtgc
     2761 cagcacacca ttattggcgt gccaggccgc gtgaagggcc tgagtggcgg agagcgcaag
     2821 cgcctggcct ttgccagcga ggccctgacc gatccccccc tgctgatttg cgacgagccg
     2881 accagcggcc tggatagctt caccgcccac tccgtggtgc aggtgctgaa gaagctgagc
     2941 cagaagggaa agacggtcat tctgacgatc caccagccca gctcggagct gttcgagctg
     3001 tttgataaga ttctgctgat ggccgaggga cgcgtggcct tcctgggaac cccaagcgag
     3061 gccgtggatt tcttcagcta tgtgggcgcc cagtgcccca cgaattacaa ccccgccgac
     3121 ttctatgtgc aagtgctggc cgtggtgcca ggacgcgaga ttgagagccg cgatcgcatt
     3181 gccaagatct gcgataactt cgccatctcc aaggtggccc gcgacatgga gcagctgctg
     3241 gccacgaaga acctggagaa gcccctggag cagcccgaga acggctatac gtataaggct
     3301 acctggttta tgcagtttcg cgccgtgctg tggcgcagct ggctgagtgt gctgaaggag
     3361 cccctgctgg tcaaggtccg cctgatccag accacgatgg tggccattct gatcggcctg
     3421 atcttcctgg gacagcagct gacgcaggtc ggagtgatga acatcaatgg agccattttt
     3481 ctgtttctga cgaacatgac gttccagaac gtgttcgcca ccatcaacgt gttcaccagc
     3541 gagctgcccg tgttcatgcg cgaggcccgc agccgcctgt accgctgcga tacctacttc
     3601 ctgggaaaga ccattgccga gctgccgctg ttcctgaccg tgcccctggt gtttaccgcc
     3661 attgcctacc ccatgattgg cctgcgtgcc ggagtgctgc atttctttaa ttgcctggcc
     3721 ctggtgaccc tggtggccaa cgtgtcgacc agcttcggct acctgatcag ctgcgcctcc
     3781 agcagcacca gcatggccct gagcgtggga ccccccgtga ttatcccctt cctgctgttc
     3841 ggcggattct ttctgaactc cggcagcgtg cccgtgtacc tgaagtggct gagctacctg
     3901 agctggtttc gctacgccaa tgagggcctg ctgatcaatc agtgggccga tgtggagccc
     3961 ggcgagatct cctgcaccag cagcaacacg acctgcccca gcagcggaaa ggtcatcctg
     4021 gagacactga atttcagtgc cgccgacctg ccactggatt atgtgggcct ggccatcctg
     4081 attgtgtcct tccgcgtgct ggcctatctg gctctgcgcc tgcgcgctcg tcgcaaggag
     4141 tgagcttgat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4201 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4261 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4321 gggggaggtg tgggaggttt tttcagtatg gcaagcgctc gacgaattaa caagcttcgc
     4381 tctatctcgc tctctatctc tctcgatcgc tcgtgctaag cttaagaccg tggagaacaa
     4441 ggagaaccgc tgtcaggagg catgcgtcga ctctagaccc gggcagctgc gcttgacacg
     4501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5281 cacctagatc cttttgtgac cagccaggac agaaatgcct cgacttcgct gctacccaag
     5341 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
     5401 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
     5461 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
     5521 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     5581 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     5641 aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     5701 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     5761 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     5821 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     5881 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     5941 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     6001 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     6061 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     6121 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     6181 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     6241 gcagtaaccg gcaaaatcgc gccgaaggat gtagctgccg actgggcaat ggagcgcctg
     6301 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     6361 gatcgtttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     6421 atcaccaagg tagtcggcaa ataatcacta gaccaatgtt acacatatat actttagatt
     6481 gatttaaaac ttcatgttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6541 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6601 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6661 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6721 atggttcacg tagtgggcca gcgccctgat agacggtttt tcgccctttg acgttggagt
     6781 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6841 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6901 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca attttaacgg
     6961 atcagcgaga gacccctgtt gacaattaat catcggcata gtatatcggc atagtataat
     7021 acgacaaggt gaggaactaa acccaggagg cagatcatga ccatgattac gccaagctat
     7081 ttaggtgaca ctatagaata ctc
//
