LOCUS       sequence-12198-e        2924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2924)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2924)
  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-J72019
FEATURES             Location/Qualifiers
     source          1..2924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             48..71
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(98..115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(177..201)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        248..352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(353..421,422..1213)
                     /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(571..590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1494..1513)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1505..2320
                     /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             1505..2320
                     /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(1578..1597)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2403..2759
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2821..2845
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgggac aatcccggtc tttaagaaca ggaattacat gtctgacgat tacaaggatg
       61 acgacgacaa gggatcctaa ctcgctcctc aggcttcctc gctcactgac tcgctgcgct
      121 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc aattcgaccc
      181 agctttcttg tacaaagtgg ttgatcggat ctcttagacg tcaggtggca cttttcgggg
      241 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      301 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      361 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      421 tcacccagaa acgctggtga aagtaaaaga tgcagaggat cagttgggtg cacgagtggg
      481 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      541 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      601 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
      661 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
      721 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
      781 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
      841 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
      901 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
      961 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1021 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1081 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1141 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1201 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1261 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatcgga tctctgaaga
     1321 ctctcgagct gcaggactca cagctcctcg gatctgctga tccttcaact cagcaaaagt
     1381 tcgatttatt caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1441 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1501 tgttatgagc catattcaac gggaaacgtc ttgctccagg ccgcgattaa attccaacat
     1561 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1621 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1681 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1741 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1801 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1861 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1921 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1981 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2041 gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt
     2101 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2161 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2221 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2281 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2341 gtaacactgg cagagcatta cgctgacttg acgggaggat ctagcagttc aacctgttga
     2401 tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct
     2461 ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac
     2521 taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc
     2581 agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc
     2641 ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag
     2701 aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat
     2761 gggaattagc catgggcccg tgcgaatcac ggatctcccg atcaagaatt gccattatca
     2821 acaagtttgt acaaaaaagc aggctccgaa ttggtatcac gaggcagaat ttcagataaa
     2881 aaaaatcctt agctttcgct aaggatctga agtggaattc atga
//
