LOCUS       sequence-12304-e        3051 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 3051)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 3051)
  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-J72038
FEATURES             Location/Qualifiers
     source          1..3051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(431..448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(510..534)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(684..703)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             695..1510
                     /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             695..1510
                     /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(768..787)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1720..2379
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1786..1805)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2400..2494
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2530..2886
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2948..2972
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctatgac cgaccgtctg acccaactcc aaatttgcct ggaccagatg accgaacagt
       61 tctgcgctac cctgaactac atcgacaaaa accacggttt cgaacgtctg accgttaacg
      121 aaccgcagat gtctgacaaa cacgctaccg ttgttccgcc ggaagagttt tctaacacta
      181 ttgacgaact ctccaccgac atcatcctga aaacccgtca gatcaacaaa ctgatcgact
      241 ctctgccagg cgttgacgtt tccgcagagg aacaactccg taaaatcgac atgctccaaa
      301 aaaagctcgt tgaagttgaa gacgaaaaaa tcgaagctat caaaaaaaaa gaaaaactgc
      361 tgcgtcacgt tgactctctg atcgaagact tcgttgacgg ttaaggatcc taactcgctc
      421 ctcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      481 cagctcactc aaaggcggta atcaattcga cccagctttc ttgtacaaag tggttgatcg
      541 gatctgctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc
      601 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
      661 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
      721 ttgctccagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
      781 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
      841 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
      901 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
      961 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
     1021 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     1081 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     1141 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     1201 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     1261 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     1321 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     1381 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     1441 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     1501 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     1561 acgggaggat ctctgaagac tctcgagctg caggactcac agctcctcgg atctttgatc
     1621 gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
     1681 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1741 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1801 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1861 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1921 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1981 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     2041 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     2101 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     2161 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     2221 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     2281 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     2341 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg
     2401 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     2461 cggttgccgc cgggcgtttt ttattggtga gaatccaagc ctcggatcta gcagttcaac
     2521 ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat gtttaacgta
     2581 ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc tctcatggct
     2641 aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa taaaattaat
     2701 ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa aaaagaatat
     2761 ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag ggatgtaacg
     2821 cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa cacttaacgg
     2881 ctgacatggg aattagccat gggcccgtgc gaatcacgga tctcccgatc aagaattgcc
     2941 attatcaaca agtttgtaca aaaaagcagg ctccgaattg gtatcacgag gcagaatttc
     3001 agataaaaaa aatccttagc tttcgctaag gatctgaagt ggaattcatg a
//
