LOCUS       sequence-12305-e        3208 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 3208)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 3208)
  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-J72039
FEATURES             Location/Qualifiers
     source          1..3208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(382..399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(461..485)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        532..636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(637..705,706..1497)
                     /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(855..874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1778..1797)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1789..2604
                     /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             1789..2604
                     /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(1862..1881)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2687..3043
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    3105..3129
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctcagct aaaatcaacg tattaaggca ctccatagcg atgaaaaaat ctaccgaaaa
       61 cgaatctacc aactaccagt acaaaatcca ggaactgcgt aaactgctga aatctctgct
      121 gctgaactac ctggaactga tcggtgttct gtctatcaac ccggacatgt acgaacgtaa
      181 agttgaaaac atccgtacca tcctggttaa catccaccac ctgctgaacg aataccgtcc
      241 gcaccagtct cgtgaatctc tgatcatgct gctggaagaa cagctggaat acaaacgtgg
      301 tgaaatccgt gaaatcgaac aggtttgcaa acaggttcac gacaaactga cctctggatc
      361 ctaactcgct cctcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      421 gcgagcggta tcagctcact caaaggcggt aatcaattcg acccagcttt cttgtacaaa
      481 gtggttgatc ggatctctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      541 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      601 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      661 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      721 gtgaaagtaa aagatgcaga ggatcagttg ggtgcacgag tgggttacat cgaactggat
      781 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      841 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
      901 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      961 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1021 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1081 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1141 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1201 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1261 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1321 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     1381 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1441 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1501 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1561 aggatctagg tgaagatcct ttttgataat cggatctctg aagactctcg agctgcagga
     1621 ctcacagctc ctcggatctg ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     1681 agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     1741 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1801 caacgggaaa cgtcttgctc caggccgcga ttaaattcca acatggatgc tgatttatat
     1861 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     1921 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1981 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2041 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
     2101 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2161 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2221 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2281 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
     2341 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2401 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2461 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2521 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2581 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2641 attacgctga cttgacggga ggatctagca gttcaacctg ttgatagtac gtactaagct
     2701 ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac
     2761 taaaccctca tggctaacgt actaagctct catggctaac gtactaagct ctcatgtttc
     2821 acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac ttaaatagcc
     2881 tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa agcttttaag
     2941 gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc ttagagcctc
     3001 tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat tagccatggg
     3061 cccgtgcgaa tcacggatct cccgatcaag aattgccatt atcaacaagt ttgtacaaaa
     3121 aagcaggctc cgaattggta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     3181 cgctaaggat ctgaagtgga attcatga
//
