LOCUS       sequence-12203-e        2928 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 2928)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2928)
  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-J72024
FEATURES             Location/Qualifiers
     source          1..2928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             52..75
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(102..119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(181..205)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        252..356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(357..425,426..1217)
                     /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(575..594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1498..1517)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1509..2324
                     /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             1509..2324
                     /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(1582..1601)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2407..2763
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2825..2849
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatcttacaa ctacacacta tctcatacgg agccctaaca tgtctgacac ctggagccac
       61 ccgcagttcg aaaaaggatc ctaactcgct cctcaggctt cctcgctcac tgactcgctg
      121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatcaattcg
      181 acccagcttt cttgtacaaa gtggttgatc ggatctctta gacgtcaggt ggcacttttc
      241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgcaga ggatcagttg ggtgcacgag
      481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      901 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat cggatctctg
     1321 aagactctcg agctgcagga ctcacagctc ctcggatctg ctgatccttc aactcagcaa
     1381 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     1441 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     1501 ggggtgttat gagccatatt caacgggaaa cgtcttgctc caggccgcga ttaaattcca
     1561 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1621 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1681 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1741 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1801 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1861 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1921 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1981 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2041 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2101 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2161 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2221 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2281 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     2341 ggttgtaaca ctggcagagc attacgctga cttgacggga ggatctagca gttcaacctg
     2401 ttgatagtac gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta
     2461 agctctcatg tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac
     2521 gtactaagct ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata
     2581 aatcagcaac ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata
     2641 aggcttttaa agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac
     2701 tgagaagccc ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg
     2761 acatgggaat tagccatggg cccgtgcgaa tcacggatct cccgatcaag aattgccatt
     2821 atcaacaagt ttgtacaaaa aagcaggctc cgaattggta tcacgaggca gaatttcaga
     2881 taaaaaaaat ccttagcttt cgctaaggat ctgaagtgga attcatga
//
