LOCUS       sequence-12205-e        2936 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 2936)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2936)
  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: pMLLKA-J72026
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             25..33
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             42..83
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(110..127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(189..213)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        260..364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(365..433,434..1225)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(583..602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1506..1525)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1517..2332
                     /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             1517..2332
                     /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(1590..1609)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2415..2771
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2833..2857
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctgaagc aatatcttca cgtataagga ggtatttcat gggtaagcct atccctaacc
       61 ctctgctggg tctcgattct acgggatcct aactcgctcc tcaggcttcc tcgctcactg
      121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      181 tcaattcgac ccagctttct tgtacaaagt ggttgatcgg atctcttaga cgtcaggtgg
      241 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      361 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      421 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgcagagg atcagttggg
      481 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      541 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      601 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      661 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      721 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      781 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      841 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      901 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      961 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1021 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1081 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1141 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1201 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1261 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatcg
     1321 gatctctgaa gactctcgag ctgcaggact cacagctcct cggatctgct gatccttcaa
     1381 ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctgatgtta
     1441 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1501 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcca ggccgcgatt
     1561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1621 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1801 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     1861 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     1921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2041 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     2101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2161 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2221 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2281 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2341 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggagg atctagcagt
     2401 tcaacctgtt gatagtacgt actaagctct catgtttcac gtactaagct ctcatgttta
     2461 acgtactaag ctctcatgtt taacgaacta aaccctcatg gctaacgtac taagctctca
     2521 tggctaacgt actaagctct catgtttcac gtactaagct ctcatgtttg aacaataaaa
     2581 ttaatataaa tcagcaactt aaatagcctc taaggtttta agttttataa gaaaaaaaag
     2641 aatatataag gcttttaaag cttttaaggt ttaacggttg tggacaacaa gccagggatg
     2701 taacgcactg agaagccctt agagcctctc aaagcaattt tgagtgacac aggaacactt
     2761 aacggctgac atgggaatta gccatgggcc cgtgcgaatc acggatctcc cgatcaagaa
     2821 ttgccattat caacaagttt gtacaaaaaa gcaggctccg aattggtatc acgaggcaga
     2881 atttcagata aaaaaaatcc ttagctttcg ctaaggatct gaagtggaat tcatga
//
