LOCUS       sequence-335017-        2129 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Modified Alpha level R6K_ origin GB Cloning vector, Conditionally 
            replicative ori, requires pir gene expression requires pir gene 
            expression requires EC100D or EC100D116 bacterial strain (DH10B 
            derived) uses blue-white screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2129)
  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 2129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pR6Kg-Alpha1spm
FEATURES             Location/Qualifiers
     source          1..2129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..74
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        128..146
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     128..145
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    153..209
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(211..230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(212..230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(237..254)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(237..253)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(246..268)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(462..481)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      569..925
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(981..1772)
                     /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"
ORIGIN
        1 gaattcgtct caggagagag acccctgttg acaattaatc atcggcatag tatatcggca
       61 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgac catgattacg
      121 ccaagctatt taggtgacac tatagaatac tcaagcttgc atgcctgcag gtcgactcta
      181 gaggatcccc gggtaccgag ctcgaattcg ccctatagtg agtcgtatta caattcactg
      241 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      301 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      361 tcccaacagt tgcgcagcct gaatggcgaa tgacgcctga tgcggtattt tctccttacg
      421 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
      481 gcatagggtc tctcgctgtc atgagacgaa ttctgacgta ccgcggccgc gtagaggatc
      541 tgttgatcag cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta
      601 agctctcatg tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac
      661 gtactaagct ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg
      721 tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt
      781 ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca
      841 acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg
      901 acacaggaac acttaacggc tgacatgtga tgaagtttta aatcaatcta aagtatatat
      961 gtgtaacatt ggtctagtga ttatttgccg actactttgg tgatctcgcc tttaacgtag
     1021 tggacaaact cttccaactg atctgcgcgc gaggccaaac gatcttcttc ttgtccaaga
     1081 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
     1141 cagtcggcag ctacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
     1201 gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc
     1261 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
     1321 tccgccgctg gacctaccaa cgcaacgcta tgttctcttg cttttgtcag caagatagcc
     1381 agatcaatgt cgatggtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
     1441 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgaaca
     1501 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
     1561 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttaccgt caccgtaacc
     1621 agcaaatcaa tatcgctgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
     1681 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
     1741 gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt agggcgactg
     1801 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     1861 gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc ataacaagcc
     1921 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     1981 cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta tgtccactgg
     2041 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttgtg tagcagcgaa
     2101 gtcgaggcat ttctgtcctg gctggtaac
//
