LOCUS       sequence-12301-e        2697 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 2697)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2697)
  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-J72035
FEATURES             Location/Qualifiers
     source          1..2697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..47
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(77..94)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(156..180)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(330..349)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             341..1156
                     /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             341..1156
                     /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(414..433)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             1366..2025
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1432..1451)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2046..2140
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2176..2532
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2594..2618
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctggtaa gcctatccct aaccctctgc tgggtctcga ttctacgtaa ggatcctaac
       61 tcgctcctca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      121 cggtatcagc tcactcaaag gcggtaatca attcgaccca gctttcttgt acaaagtggt
      181 tgatcggatc tgctgatcct tcaactcagc aaaagttcga tttattcaac aaagccacgt
      241 tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
      301 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
      361 aacgtcttgc tccaggccgc gattaaattc caacatggat gctgatttat atgggtataa
      421 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
      481 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
      541 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
      601 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
      661 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
      721 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
      781 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
      841 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc
      901 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
      961 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1021 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1081 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1141 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     1201 gacttgacgg gaggatctct gaagactctc gagctgcagg actcacagct cctcggatct
     1261 ttgatcgggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1321 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1381 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1441 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1501 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1561 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     1621 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     1681 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     1741 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     1801 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     1861 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1921 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1981 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga tatcgagctc
     2041 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     2101 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcctcg gatctagcag
     2161 ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     2221 aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc
     2281 atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa
     2341 attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa
     2401 gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat
     2461 gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca caggaacact
     2521 taacggctga catgggaatt agccatgggc ccgtgcgaat cacggatctc ccgatcaaga
     2581 attgccatta tcaacaagtt tgtacaaaaa agcaggctcc gaattggtat cacgaggcag
     2641 aatttcagat aaaaaaaatc cttagctttc gctaaggatc tgaagtggaa ttcatga
//
