LOCUS       sequence-12306-e        3284 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3284)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 3284)
  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"
COMMENT     Sequence Label: pMLLKA-J72040
FEATURES             Location/Qualifiers
     source          1..3284
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(458..475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(537..561)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        608..712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(713..781,782..1573)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(931..950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1854..1873)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1865..2680
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1865..2680
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1938..1957)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2763..3119
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    3181..3205
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgaata ccctactaag gacattcgcc agaaaatgac cgaccgtctg acccaactcc
       61 aaatttgcct ggaccagatg accgaacagt tctgcgctac cctgaactac atcgacaaaa
      121 accacggttt cgaacgtctg accgttaacg aaccgcagat gtctgacaaa cacgctaccg
      181 ttgttccgcc ggaagagttt tctaacacta ttgacgaact ctccaccgac atcatcctga
      241 aaacccgtca gatcaacaaa ctgatcgact ctctgccagg cgttgacgtt tccgcagagg
      301 aacaactccg taaaatcgac atgctccaaa aaaagctcgt tgaagttgaa gacgaaaaaa
      361 tcgaagctat caaaaaaaaa gaaaaactgc tgcgtcacgt tgactctctg atcgaagact
      421 tcgttgacgg tggatcctaa ctcgctcctc aggcttcctc gctcactgac tcgctgcgct
      481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc aattcgaccc
      541 agctttcttg tacaaagtgg ttgatcggat ctcttagacg tcaggtggca cttttcgggg
      601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      781 tcacccagaa acgctggtga aagtaaaaga tgcagaggat cagttgggtg cacgagtggg
      841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatcgga tctctgaaga
     1681 ctctcgagct gcaggactca cagctcctcg gatctgctga tccttcaact cagcaaaagt
     1741 tcgatttatt caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1801 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1861 tgttatgagc catattcaac gggaaacgtc ttgctccagg ccgcgattaa attccaacat
     1921 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1981 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2041 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2101 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2161 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2221 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2281 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2341 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2401 gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt
     2461 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2521 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2581 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2641 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2701 gtaacactgg cagagcatta cgctgacttg acgggaggat ctagcagttc aacctgttga
     2761 tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct
     2821 ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac
     2881 taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc
     2941 agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc
     3001 ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag
     3061 aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat
     3121 gggaattagc catgggcccg tgcgaatcac ggatctcccg atcaagaatt gccattatca
     3181 acaagtttgt acaaaaaagc aggctccgaa ttggtatcac gaggcagaat ttcagataaa
     3241 aaaaatcctt agctttcgct aaggatctga agtggaattc atga
//
