LOCUS       sequence-12202-e        2939 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 2939)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2939)
  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-J72023
FEATURES             Location/Qualifiers
     source          1..2939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             42..86
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(113..130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(192..216)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        263..367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(368..436,437..1228)
                     /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(586..605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1509..1528)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1520..2335
                     /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             1520..2335
                     /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(1593..1612)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2418..2774
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2836..2860
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgaaaa ggaacccacg ccaagtccag gacacgccat gaaagaaacc gctgctgcta
       61 aattcgaacg tcagcacatg gattccggat cctaactcgc tcctcaggct tcctcgctca
      121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      181 taatcaattc gacccagctt tcttgtacaa agtggttgat cggatctctt agacgtcagg
      241 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      301 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      361 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      421 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgcag aggatcagtt
      481 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      541 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      601 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      661 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      721 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      781 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      841 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      901 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      961 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1021 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1081 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1141 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1201 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1261 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1321 tcggatctct gaagactctc gagctgcagg actcacagct cctcggatct gctgatcctt
     1381 caactcagca aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg
     1441 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1501 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg
     1561 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1621 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     1681 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1741 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1801 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     1861 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1921 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     1981 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2041 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     2101 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2161 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2221 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2281 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2341 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg aggatctagc
     2401 agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt
     2461 ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc
     2521 tcatggctaa cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata
     2581 aaattaatat aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa
     2641 aagaatatat aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg
     2701 atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca
     2761 cttaacggct gacatgggaa ttagccatgg gcccgtgcga atcacggatc tcccgatcaa
     2821 gaattgccat tatcaacaag tttgtacaaa aaagcaggct ccgaattggt atcacgaggc
     2881 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tctgaagtgg aattcatga
//
