LOCUS       sequence-12293-e        2694 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 2694)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2694)
  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-J72030
FEATURES             Location/Qualifiers
     source          1..2694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(74..91)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(153..177)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(327..346)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             338..1153
                     /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             338..1153
                     /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(411..430)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1363..2022
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1429..1448)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2043..2137
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2173..2529
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2591..2615
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctggtgc tccggttcca tatccagatc cactggaacc acgttaagga tcctaactcg
       61 ctcctcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      121 tatcagctca ctcaaaggcg gtaatcaatt cgacccagct ttcttgtaca aagtggttga
      181 tcggatctgc tgatccttca actcagcaaa agttcgattt attcaacaaa gccacgttgt
      241 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
      301 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
      361 gtcttgctcc aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
      421 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
      481 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
      541 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
      601 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
      661 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
      721 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
      781 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
      841 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
      901 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
      961 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     1021 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     1081 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     1141 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     1201 ttgacgggag gatctctgaa gactctcgag ctgcaggact cacagctcct cggatctttg
     1261 atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     1321 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     1381 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1441 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1501 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1561 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1621 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1681 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1741 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     1801 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     1861 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     1921 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     1981 gaattacaac agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct
     2041 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     2101 gctcggttgc cgccgggcgt tttttattgg tgagaatcca agcctcggat ctagcagttc
     2161 aacctgttga tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac
     2221 gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg
     2281 gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt
     2341 aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa
     2401 tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta
     2461 acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa
     2521 cggctgacat gggaattagc catgggcccg tgcgaatcac ggatctcccg atcaagaatt
     2581 gccattatca acaagtttgt acaaaaaagc aggctccgaa ttggtatcac gaggcagaat
     2641 ttcagataaa aaaaatcctt agctttcgct aaggatctga agtggaattc atga
//
