LOCUS       sequence-12298-e        2700 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 2700)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2700)
  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-J72032
FEATURES             Location/Qualifiers
     source          1..2700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..50
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(80..97)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(159..183)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(333..352)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             344..1159
                     /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             344..1159
                     /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(417..436)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1369..2028
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1435..1454)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2049..2143
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2179..2535
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2597..2621
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctaaaga aaccgctgct gctaaattcg aacgtcagca catggattcc taaggatcct
       61 aactcgctcc tcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      121 gagcggtatc agctcactca aaggcggtaa tcaattcgac ccagctttct tgtacaaagt
      181 ggttgatcgg atctgctgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca
      241 cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
      301 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
      361 ggaaacgtct tgctccaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
      421 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
      481 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
      541 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
      601 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
      661 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
      721 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
      781 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
      841 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
      901 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
      961 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1021 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1081 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1141 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     1201 gctgacttga cgggaggatc tctgaagact ctcgagctgc aggactcaca gctcctcgga
     1261 tctttgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     1321 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1381 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1441 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1501 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1561 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1621 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1681 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1741 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1801 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1861 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1921 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1981 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag
     2041 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2101 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagcc tcggatctag
     2161 cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg
     2221 tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac gtactaagct
     2281 ctcatggcta acgtactaag ctctcatgtt tcacgtacta agctctcatg tttgaacaat
     2341 aaaattaata taaatcagca acttaaatag cctctaaggt tttaagtttt ataagaaaaa
     2401 aaagaatata taaggctttt aaagctttta aggtttaacg gttgtggaca acaagccagg
     2461 gatgtaacgc actgagaagc ccttagagcc tctcaaagca attttgagtg acacaggaac
     2521 acttaacggc tgacatggga attagccatg ggcccgtgcg aatcacggat ctcccgatca
     2581 agaattgcca ttatcaacaa gtttgtacaa aaaagcaggc tccgaattgg tatcacgagg
     2641 cagaatttca gataaaaaaa atccttagct ttcgctaagg atctgaagtg gaattcatga
//
