LOCUS       sequence-12294-e        2679 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 2679)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2679)
  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: pMLLCK-K112603
FEATURES             Location/Qualifiers
     source          1..2679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..29
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(59..76)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(138..162)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(312..331)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             323..1138
                     /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             323..1138
                     /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(396..415)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1348..2007
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1414..1433)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2028..2122
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2158..2514
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2576..2600
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgacta caaggatgac gacgacaagt aaggatccta actcgctcct caggcttcct
       61 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      121 aggcggtaat caattcgacc cagctttctt gtacaaagtg gttgatcgga tctgctgatc
      181 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg
      241 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
      301 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc
      361 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      421 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      481 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      541 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      601 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
      661 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      721 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      781 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
      841 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
      901 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
      961 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1021 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1081 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1141 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggaggatct
     1201 ctgaagactc tcgagctgca ggactcacag ctcctcggat ctttgatcgg gcacgtaaga
     1261 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1321 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1381 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1441 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1501 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1561 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1621 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1681 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1741 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1801 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1861 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1921 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1981 tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat
     2041 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     2101 ggcgtttttt attggtgaga atccaagcct cggatctagc agttcaacct gttgatagta
     2161 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat
     2221 gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc
     2281 tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa
     2341 cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta
     2401 aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc
     2461 cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa
     2521 ttagccatgg gcccgtgcga atcacggatc tcccgatcaa gaattgccat tatcaacaag
     2581 tttgtacaaa aaagcaggct ccgaattggt atcacgaggc agaatttcag ataaaaaaaa
     2641 tccttagctt tcgctaagga tctgaagtgg aattcatga
//
