LOCUS       sequence-323401-        7099 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 7099)
  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 7099)
  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..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(511..1302)
                     /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      1770..2358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2259..2278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2669..2803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2706..2725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2760..2779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(6017..6468)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(6048..6069)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    6496..6565
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     6622..6641
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6835..6857
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6849..6866
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6850..6866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6873..6892
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6873..6891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6894..6950
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(6957..6975)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6958..6975)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(7029..7076)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaca tgaagtttta aatcaatcta
      481 aagtatatat gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc
      541 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaaac gatcttcttc
      601 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
      661 ctccattgcc cagtcggcag ctacatcctt cggcgcgatt ttgccggtta ctgcgctgta
      721 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
      781 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
      841 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
      901 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
      961 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     1021 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     1081 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     1141 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     1201 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     1261 tgatagttga gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt
     1321 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     1381 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
     1441 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     1501 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
     1561 tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     1621 tagcagcgaa gtcgaggcat ttctgtcctg gctggtcaca aaaggatcta ggtgaagatc
     1681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtgtc
     2461 aagcgcagct gcccgggtct agagtcgacg catgcctcct gacgttctcc ttgttctcca
     2521 cggtcttaag cttagcacga gcgatcgaga gagatagaga gcgagataga gcgaagcttg
     2581 ttaattcgtc gagcgcttgc catactgaaa aaacctccca cacctccccc tgaacctgaa
     2641 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2701 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2761 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaagctca ctccttgcga
     2821 cgagcgcgca ggcgcagagc cagataggcc agcacgcgga aggacacaat caggatggcc
     2881 aggcccacat aatccagtgg caggtcggcg gcactgaaat tcagtgtctc caggatgacc
     2941 tttccgctgc tggggcaggt cgtgttgctg ctggtgcagg agatctcgcc gggctccaca
     3001 tcggcccact gattgatcag caggccctca ttggcgtagc gaaaccagct caggtagctc
     3061 agccacttca ggtacacggg cacgctgccg gagttcagaa agaatccgcc gaacagcagg
     3121 aaggggataa tcacgggggg tcccacgctc agggccatgc tggtgctgct ggaggcgcag
     3181 ctgatcaggt agccgaagct ggtcgacacg ttggccacca gggtcaccag ggccaggcaa
     3241 ttaaagaaat gcagcactcc ggcacgcagg ccaatcatgg ggtaggcaat ggcggtaaac
     3301 accaggggca cggtcaggaa cagcggcagc tcggcaatgg tctttcccag gaagtaggta
     3361 tcgcagcggt acaggcggct gcgggcctcg cgcatgaaca cgggcagctc gctggtgaac
     3421 acgttgatgg tggcgaacac gttctggaac gtcatgttcg tcagaaacag aaaaatggct
     3481 ccattgatgt tcatcactcc gacctgcgtc agctgctgtc ccaggaagat caggccgatc
     3541 agaatggcca ccatcgtggt ctggatcagg cggaccttga ccagcagggg ctccttcagc
     3601 acactcagcc agctgcgcca cagcacggcg cgaaactgca taaaccaggt agccttatac
     3661 gtatagccgt tctcgggctg ctccaggggc ttctccaggt tcttcgtggc cagcagctgc
     3721 tccatgtcgc gggccacctt ggagatggcg aagttatcgc agatcttggc aatgcgatcg
     3781 cggctctcaa tctcgcgtcc tggcaccacg gccagcactt gcacatagaa gtcggcgggg
     3841 ttgtaattcg tggggcactg ggcgcccaca tagctgaaga aatccacggc ctcgcttggg
     3901 gttcccagga aggccacgcg tccctcggcc atcagcagaa tcttatcaaa cagctcgaac
     3961 agctccgagc tgggctggtg gatcgtcaga atgaccgtct ttcccttctg gctcagcttc
     4021 ttcagcacct gcaccacgga gtgggcggtg aagctatcca ggccgctggt cggctcgtcg
     4081 caaatcagca gggggggatc ggtcagggcc tcgctggcaa aggccaggcg cttgcgctct
     4141 ccgccactca ggcccttcac gcggcctggc acgccaataa tggtgtgctg gcacttgctc
     4201 aggctcagct cctggatgac ctggtccacg cgggccacgc gctgacggta tgtcaggtgg
     4261 cggggcatgc ggaccatggc ctgaaagatc agatgctcgc gggctgtcag gctgccgatg
     4321 aacagatcgt cctgctgcac gtaggcgcag cgggcctgca tctccttggc atccacgggc
     4381 tggccgttca gcaggcgcat gccgcttggg gacacctgaa tgccctgggg gctgcggaag
     4441 gccagggcgt tcagcagcgt ggtctttccg gcgccactac ttcccatcac ggccagcagc
     4501 tcgccggggt aggccactcc gcacacattc ttcagcaggt gcttgcgtgg ggctgggata
     4561 tggcgctcat tgcagaacag tccgcgggtg cggttcacca gctgacgcca gccactgcct
     4621 ggctggttca cggcgccgaa gatatccatg ttatgccagg cgtacgtcag attctcggcc
     4681 agctgtccgg atccgcttcc gctatcctct ggggggctgg ggggcagcag ggttccgtaa
     4741 ttcttggcct ggccaaatcc ctgattgatg cacgactggc tggcggctcc gctatcgccg
     4801 ttgttcaggt gctcggcgct tgggtgcttg gagcctccgc ggatcagcag ctcctggtcc
     4861 tcctgtccca tttggtttgg tatgcgtctt gtgattcaaa gttggcttat tcaaaggata
     4921 ttaacgaagg cagcggcacg tctgctcggc tcgaacattc attcacaact gatgctctca
     4981 gccaccccgc gcccttttat accgctgcgc tcgatccccg ggcgagctcg ggaggggcat
     5041 gacgtaccta cctcgactgc tactcgactt aaaggcattt caagggtttc cactgggctc
     5101 gacttaaagg catttcaagg gtttccactg ggctcgactt aaaggcattt caagggtttc
     5161 cactgggctc gacttaaagg catttcaagg gtttccactg ggctcgactt aaaggcattt
     5221 caagggtttc cactgggctc gacagtcagg tgctagtcgt acgctcctga cagcggaaca
     5281 aacgacccaa cacccgtgcg ttttattctg tctttttatt gccaagctta ggctcctggc
     5341 ttacgggtca tacaccaggt gcgtgggccc tcgggcacct ccacatcggc cgtcacggtg
     5401 aagcccagac gctcgtagaa gggcagattg cgtggggcac tggtttccag gaaggctggc
     5461 actccagcac gctcggcagc ttccactcct ggcagcacca cggcactgcc caggcccttg
     5521 ccctggtggt cggggctcac gcccacggtg gccaggaacc agacgggctc ctttggacgg
     5581 tgtggggcca gcaggccctc catctgctgc tgggcagcca gacgggagcc actcagctcg
     5641 gccatacggg gtccaatctc ggcgaacacg gctccagcct ccacgctctc gggggtggtc
     5701 cacacggcca cggcagctcc atcatcggcc acccagacct tgccgatatc caggcccacg
     5761 cgtgtcagga acagctcctg gagctcggtc acacgctcaa tgtgacggtc tgggtccacg
     5821 gtgtgacggg tggctgggta atcggcgaag gcagcagcca gggttcgcac ggcacggggc
     5881 acatcatcac gggtggccag acgcaccgtg ggcttatact cggtcatttt gttttcctcc
     5941 ttatgttaag cttactcagt tattatatca taaatatctg tgtcaagaat aaactcccac
     6001 atggattcga gtattgagag ttctcttctt gtattcaata attacttctt ggcagatttc
     6061 agtagttgca gttgatttac ttggttgctg gttactttta attgattcac tttaacttgc
     6121 actttactgc agattgttta gcttgttcag ctgcgcttgt ttatttgctt agctttcgct
     6181 tagcgacgtg ttcactttgc ttgtttgaat tgaattgtcg ctccgtagac gaagcgcctc
     6241 tatttatact ccggcgctct tttcgcgaac attcgaggcg cgctctctcg aaccaacgag
     6301 agcagtatgc cgtttactgt gtgacagagt gagagagcat tagtgcagag agggagcgac
     6361 ccaaaaagaa aagagagaat aacgaataac ggccagagaa atttctcgag ttttctttct
     6421 gccaaacaaa tgacctacca caataaccag tttgttttgg gattctagct cctgacagcg
     6481 catcatgatg gaccagatgg gtgaggtgga gtacgcgccc ggggagccca agggcacgcc
     6541 ctggcacccg caccgcggct tcgagacagt gacctacatc gtcgaccaat tcctccctgg
     6601 gatccggaga gagaccctat gcggcatcag agcagattgt actgagagtg caccatatgc
     6661 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgt cattcgccat
     6721 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6781 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6841 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattcg
     6901 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt gagtattcta
     6961 tagtgtcacc taaatagctt ggcgtaatca tggtcatgat ctgcctcctg ggtttagttc
     7021 ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt gtcaacaggg
     7081 gtctctcgct gatccgtta
//
