LOCUS       sequence-12195-e        2915 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 2915)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2915)
  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-J72016
FEATURES             Location/Qualifiers
     source          1..2915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             45..62
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(89..106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(168..192)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        239..343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(344..412,413..1204)
                     /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(562..581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1496..2311
                     /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             1496..2311
                     /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(1569..1588)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2394..2750
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2812..2836
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctctaca acacctagca tctaagaagt tacatatgga ttctcaccat catcaccatc
       61 acggatccta actcgctcct caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
      121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat caattcgacc cagctttctt
      181 gtacaaagtg gttgatcgga tctcttagac gtcaggtggc acttttcggg gaaatgtgcg
      241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      421 aacgctggtg aaagtaaaag atgcagagga tcagttgggt gcacgagtgg gttacatcga
      481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      541 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      721 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
      901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1021 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1201 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1261 atttaaaagg atctaggtga agatcctttt tgataatcgg atctctgaag actctcgagc
     1321 tgcaggactc acagctcctc ggatctgctg atccttcaac tcagcaaaag ttcgatttat
     1381 tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1441 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1501 ccatattcaa cgggaaacgt cttgctccag gccgcgatta aattccaaca tggatgctga
     1561 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1621 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1681 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1741 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1801 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1861 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1921 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1981 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2041 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2101 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2161 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2221 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2281 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2341 gcagagcatt acgctgactt gacgggagga tctagcagtt caacctgttg atagtacgta
     2401 ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt
     2461 aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc
     2521 atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta
     2581 aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc
     2641 ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta
     2701 gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca tgggaattag
     2761 ccatgggccc gtgcgaatca cggatctccc gatcaagaat tgccattatc aacaagtttg
     2821 tacaaaaaag caggctccga attggtatca cgaggcagaa tttcagataa aaaaaatcct
     2881 tagctttcgc taaggatctg aagtggaatt catga
//
