LOCUS       sequence-12197-e        2935 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 2935)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2935)
  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-J72018
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(109..126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(188..212)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        259..363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(364..432,433..1224)
                     /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(582..601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1505..1524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1516..2331
                     /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             1516..2331
                     /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(1589..1608)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2414..2770
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2832..2856
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctcacat agcggcgcta aataataggg aggcgattca atgggtgctc cggttccata
       61 tccagatcca ctggaaccac gtggatccta actcgctcct caggcttcct cgctcactga
      121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      181 caattcgacc cagctttctt gtacaaagtg gttgatcgga tctcttagac gtcaggtggc
      241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      301 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      361 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      421 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgcagagga tcagttgggt
      481 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      541 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      601 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      661 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      721 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      781 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      841 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      901 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      961 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1021 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1081 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1141 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1201 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1261 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatcgg
     1321 atctctgaag actctcgagc tgcaggactc acagctcctc ggatctgctg atccttcaac
     1381 tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac
     1441 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1501 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctccag gccgcgatta
     1561 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1621 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1681 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1741 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1801 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
     1861 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1921 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1981 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2041 gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact
     2101 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2161 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2221 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2281 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2341 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggagga tctagcagtt
     2401 caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa
     2461 cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat
     2521 ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat
     2581 taatataaat cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga
     2641 atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt
     2701 aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta
     2761 acggctgaca tgggaattag ccatgggccc gtgcgaatca cggatctccc gatcaagaat
     2821 tgccattatc aacaagtttg tacaaaaaag caggctccga attggtatca cgaggcagaa
     2881 tttcagataa aaaaaatcct tagctttcgc taaggatctg aagtggaatt catga
//
