LOCUS       sequence-12299-e        2682 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 2682)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2682)
  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-J72033
FEATURES             Location/Qualifiers
     source          1..2682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..32
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(62..79)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(141..165)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(315..334)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             326..1141
                     /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             326..1141
                     /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(399..418)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1351..2010
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1417..1436)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2031..2125
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2161..2517
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2579..2603
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctacctg gagccacccg cagttcgaaa aataaggatc ctaactcgct cctcaggctt
       61 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      121 caaaggcggt aatcaattcg acccagcttt cttgtacaaa gtggttgatc ggatctgctg
      181 atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct
      241 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
      301 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctccag
      361 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
      421 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
      481 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
      541 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
      601 tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga
      661 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
      721 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
      781 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
      841 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
      901 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
      961 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     1021 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     1081 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     1141 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggagga
     1201 tctctgaaga ctctcgagct gcaggactca cagctcctcg gatctttgat cgggcacgta
     1261 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     1321 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     1381 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     1441 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     1501 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     1561 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     1621 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     1681 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     1741 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     1801 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     1861 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     1921 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     1981 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
     2041 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     2101 ccgggcgttt tttattggtg agaatccaag cctcggatct agcagttcaa cctgttgata
     2161 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct
     2221 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc taacgtacta
     2281 agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa tataaatcag
     2341 caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata tataaggctt
     2401 ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac gcactgagaa
     2461 gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg gctgacatgg
     2521 gaattagcca tgggcccgtg cgaatcacgg atctcccgat caagaattgc cattatcaac
     2581 aagtttgtac aaaaaagcag gctccgaatt ggtatcacga ggcagaattt cagataaaaa
     2641 aaatccttag ctttcgctaa ggatctgaag tggaattcat ga
//
