LOCUS       sequence-12300-e        2688 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 2688)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2688)
  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-J72034
FEATURES             Location/Qualifiers
     source          1..2688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(68..85)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(147..171)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(321..340)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             332..1147
                     /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             332..1147
                     /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(405..424)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1357..2016
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1423..1442)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2037..2131
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2167..2523
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2585..2609
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctatggc ttctatgact ggcggtcagc aaatgggtta aggatcctaa ctcgctcctc
       61 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      121 ctcactcaaa ggcggtaatc aattcgaccc agctttcttg tacaaagtgg ttgatcggat
      181 ctgctgatcc ttcaactcag caaaagttcg atttattcaa caaagccacg ttgtgtctca
      241 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
      301 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
      361 ctccaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      421 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
      481 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
      541 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
      601 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
      661 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
      721 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
      781 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
      841 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
      901 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
      961 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1021 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1081 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1141 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1201 ggaggatctc tgaagactct cgagctgcag gactcacagc tcctcggatc tttgatcggg
     1261 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1321 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1381 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1441 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1501 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     1561 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     1621 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1681 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1741 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1801 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1861 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1921 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     1981 caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct cgcttggact
     2041 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     2101 ttgccgccgg gcgtttttta ttggtgagaa tccaagcctc ggatctagca gttcaacctg
     2161 ttgatagtac gtactaagct ctcatgtttc acgtactaag ctctcatgtt taacgtacta
     2221 agctctcatg tttaacgaac taaaccctca tggctaacgt actaagctct catggctaac
     2281 gtactaagct ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata
     2341 aatcagcaac ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata
     2401 aggcttttaa agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac
     2461 tgagaagccc ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg
     2521 acatgggaat tagccatggg cccgtgcgaa tcacggatct cccgatcaag aattgccatt
     2581 atcaacaagt ttgtacaaaa aagcaggctc cgaattggta tcacgaggca gaatttcaga
     2641 taaaaaaaat ccttagcttt cgctaaggat ctgaagtgga attcatga
//
