LOCUS       sequence-12295-e        2685 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 2685)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2685)
  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-K112507
FEATURES             Location/Qualifiers
     source          1..2685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..32
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(65..82)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(144..168)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(318..337)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             329..1144
                     /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             329..1144
                     /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(402..421)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1354..2013
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1420..1439)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2034..2128
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2164..2520
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2582..2606
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatcttaccc atacgacgtc ccagactacg ctgggtaagg atcctaactc gctcctcagg
       61 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      121 actcaaaggc ggtaatcaat tcgacccagc tttcttgtac aaagtggttg atcggatctg
      181 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg tgtctcaaaa
      241 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      301 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
      361 caggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
      421 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
      481 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
      541 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
      601 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
      661 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
      721 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
      781 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
      841 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
      901 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
      961 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1021 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1081 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     1141 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     1201 ggatctctga agactctcga gctgcaggac tcacagctcc tcggatcttt gatcgggcac
     1261 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     1321 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     1381 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     1441 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     1501 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     1561 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     1621 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     1681 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     1741 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     1801 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     1861 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     1921 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     1981 cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct
     2041 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     2101 ccgccgggcg ttttttattg gtgagaatcc aagcctcgga tctagcagtt caacctgttg
     2161 atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa cgtactaagc
     2221 tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat ggctaacgta
     2281 ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat taatataaat
     2341 cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga atatataagg
     2401 cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt aacgcactga
     2461 gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta acggctgaca
     2521 tgggaattag ccatgggccc gtgcgaatca cggatctccc gatcaagaat tgccattatc
     2581 aacaagtttg tacaaaaaag caggctccga attggtatca cgaggcagaa tttcagataa
     2641 aaaaaatcct tagctttcgc taaggatctg aagtggaatt catga
//
