LOCUS       sequence-12201-e        2929 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 2929)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2929)
  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-J72022
FEATURES             Location/Qualifiers
     source          1..2929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             47..76
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(103..120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(182..206)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        253..357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(358..426,427..1218)
                     /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(576..595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1499..1518)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1510..2325
                     /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             1510..2325
                     /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(1583..1602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2408..2764
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2826..2850
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctacgca atatcacacg gaaccggagg gctaactcgc atgtctgaac aaaaactcat
       61 ctcagaagag gatctgggat cctaactcgc tcctcaggct tcctcgctca ctgactcgct
      121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatcaattc
      181 gacccagctt tcttgtacaa agtggttgat cggatctctt agacgtcagg tggcactttt
      241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      361 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      421 tttgctcacc cagaaacgct ggtgaaagta aaagatgcag aggatcagtt gggtgcacga
      481 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      541 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      601 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      661 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      721 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      781 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      841 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      901 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      961 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1021 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1081 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1141 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1201 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tcggatctct
     1321 gaagactctc gagctgcagg actcacagct cctcggatct gctgatcctt caactcagca
     1381 aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca
     1441 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1501 aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg attaaattcc
     1561 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1621 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1681 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1741 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1801 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1861 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1921 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1981 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2041 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     2101 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2161 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2221 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2281 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     2341 tggttgtaac actggcagag cattacgctg acttgacggg aggatctagc agttcaacct
     2401 gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact
     2461 aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa
     2521 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat
     2581 aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat
     2641 aaggctttta aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca
     2701 ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct
     2761 gacatgggaa ttagccatgg gcccgtgcga atcacggatc tcccgatcaa gaattgccat
     2821 tatcaacaag tttgtacaaa aaagcaggct ccgaattggt atcacgaggc agaatttcag
     2881 ataaaaaaaa tccttagctt tcgctaagga tctgaagtgg aattcatga
//
