LOCUS       sequence-12303-e        2970 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 2970)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2970)
  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-J72037
FEATURES             Location/Qualifiers
     source          1..2970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(350..367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(429..453)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(603..622)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             614..1429
                     /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             614..1429
                     /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(687..706)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1639..2298
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1705..1724)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2319..2413
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2449..2805
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2867..2891
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctatgaa aaaatctacc gaaaacgaat ctaccaacta ccagtacaaa atccaggaac
       61 tgcgtaaact gctgaaatct ctgctgctga actacctgga actgatcggt gttctgtcta
      121 tcaacccgga catgtacgaa cgtaaagttg aaaacatccg taccatcctg gttaacatcc
      181 accacctgct gaacgaatac cgtccgcacc agtctcgtga atctctgatc atgctgctgg
      241 aagaacagct ggaatacaaa cgtggtgaaa tccgtgaaat cgaacaggtt tgcaaacagg
      301 ttcacgacaa actgacctct taaggatcct aactcgctcc tcaggcttcc tcgctcactg
      361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      421 tcaattcgac ccagctttct tgtacaaagt ggttgatcgg atctgctgat ccttcaactc
      481 agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat
      541 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
      601 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctccaggc cgcgattaaa
      661 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
      721 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
      781 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
      841 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
      901 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
      961 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1021 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1081 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1141 acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag tcgtcactca
     1201 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1261 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1321 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1381 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     1441 taattggttg taacactggc agagcattac gctgacttga cgggaggatc tctgaagact
     1501 ctcgagctgc aggactcaca gctcctcgga tctttgatcg ggcacgtaag aggttccaac
     1561 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     1621 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1681 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1741 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1801 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
     1861 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     1921 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     1981 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2041 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     2101 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     2161 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     2221 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     2281 tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt
     2341 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     2401 tattggtgag aatccaagcc tcggatctag cagttcaacc tgttgatagt acgtactaag
     2461 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     2521 actaaaccct catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt
     2581 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag
     2641 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta
     2701 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc
     2761 tctcaaagca attttgagtg acacaggaac acttaacggc tgacatggga attagccatg
     2821 ggcccgtgcg aatcacggat ctcccgatca agaattgcca ttatcaacaa gtttgtacaa
     2881 aaaagcaggc tccgaattgg tatcacgagg cagaatttca gataaaaaaa atccttagct
     2941 ttcgctaagg atctgaagtg gaattcatga
//
