LOCUS       sequence-12204-e        2925 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 2925)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2925)
  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-J72025
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(99..116)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(178..202)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        249..353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(354..422,423..1214)
                     /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(572..591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1495..1514)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1506..2321
                     /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             1506..2321
                     /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(1579..1598)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2404..2760
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2822..2846
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgtacc aaattaaaaa aatcaaaacg gggtaagata tggcttctat gactggcggt
       61 cagcaaatgg gtggatccta actcgctcct caggcttcct cgctcactga ctcgctgcgc
      121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat caattcgacc
      181 cagctttctt gtacaaagtg gttgatcgga tctcttagac gtcaggtggc acttttcggg
      241 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      301 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      361 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      421 ctcacccaga aacgctggtg aaagtaaaag atgcagagga tcagttgggt gcacgagtgg
      481 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      541 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      601 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      661 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      721 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      781 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      841 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      901 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      961 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1021 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1081 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1141 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1201 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1261 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatcgg atctctgaag
     1321 actctcgagc tgcaggactc acagctcctc ggatctgctg atccttcaac tcagcaaaag
     1381 ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     1441 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1501 gtgttatgag ccatattcaa cgggaaacgt cttgctccag gccgcgatta aattccaaca
     1561 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1621 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1681 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1741 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1801 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1861 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1921 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1981 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2041 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     2101 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2161 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2221 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2281 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2341 tgtaacactg gcagagcatt acgctgactt gacgggagga tctagcagtt caacctgttg
     2401 atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc
     2461 tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta
     2521 ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat
     2581 cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg
     2641 cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga
     2701 gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca
     2761 tgggaattag ccatgggccc gtgcgaatca cggatctccc gatcaagaat tgccattatc
     2821 aacaagtttg tacaaaaaag caggctccga attggtatca cgaggcagaa tttcagataa
     2881 aaaaaatcct tagctttcgc taaggatctg aagtggaatt catga
//
