LOCUS       sequence-12291-e        2673 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 2673)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2673)
  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-J72028
FEATURES             Location/Qualifiers
     source          1..2673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..23
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(53..70)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(132..156)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(306..325)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             317..1132
                     /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             317..1132
                     /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(390..409)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1342..2001
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1408..1427)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2022..2116
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2152..2508
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2570..2594
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctcacca tcatcaccat cactaaggat cctaactcgc tcctcaggct tcctcgctca
       61 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      121 taatcaattc gacccagctt tcttgtacaa agtggttgat cggatctgct gatccttcaa
      181 ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctgatgtta
      241 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
      301 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcca ggccgcgatt
      361 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
      421 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
      481 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
      541 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
      601 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
      661 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
      721 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
      781 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
      841 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
      901 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
      961 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     1021 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     1081 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     1141 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggagg atctctgaag
     1201 actctcgagc tgcaggactc acagctcctc ggatctttga tcgggcacgt aagaggttcc
     1261 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     1321 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     1381 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     1441 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     1501 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
     1561 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     1621 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     1681 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     1741 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     1801 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     1861 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     1921 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     1981 gagtggcagg gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc
     2041 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     2101 ttttattggt gagaatccaa gcctcggatc tagcagttca acctgttgat agtacgtact
     2161 aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa
     2221 cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat
     2281 gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa
     2341 tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt
     2401 ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga
     2461 gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacatg ggaattagcc
     2521 atgggcccgt gcgaatcacg gatctcccga tcaagaattg ccattatcaa caagtttgta
     2581 caaaaaagca ggctccgaat tggtatcacg aggcagaatt tcagataaaa aaaatcctta
     2641 gctttcgcta aggatctgaa gtggaattca tga
//
