LOCUS       sequence-12200-e        2932 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 2932)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2932)
  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-J72021
FEATURES             Location/Qualifiers
     source          1..2932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..76
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     primer_bind     complement(106..123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(185..209)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        256..360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(361..429,430..1221)
                     /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(579..598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1502..1521)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1513..2328
                     /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             1513..2328
                     /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(1586..1605)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2411..2767
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2829..2853
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatctcgaac acccatatca atattaaagg gaaggtaagc atgcagccgg agctcgcacc
       61 agaagacccg gaagactgcg gatcctaact cgctcctcag gcttcctcgc tcactgactc
      121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatcaa
      181 ttcgacccag ctttcttgta caaagtggtt gatcggatct cttagacgtc aggtggcact
      241 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      301 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      361 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      421 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg cagaggatca gttgggtgca
      481 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      541 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      601 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      661 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      721 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      781 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      841 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      901 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      961 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1021 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1081 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1141 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1201 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatcggatc
     1321 tctgaagact ctcgagctgc aggactcaca gctcctcgga tctgctgatc cttcaactca
     1381 gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt
     1441 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1501 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc gcgattaaat
     1561 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1621 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1681 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1741 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1801 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     1861 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1921 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1981 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2041 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     2101 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2161 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2221 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2281 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2341 aattggttgt aacactggca gagcattacg ctgacttgac gggaggatct agcagttcaa
     2401 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     2461 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     2521 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     2581 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     2641 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     2701 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     2761 gctgacatgg gaattagcca tgggcccgtg cgaatcacgg atctcccgat caagaattgc
     2821 cattatcaac aagtttgtac aaaaaagcag gctccgaatt ggtatcacga ggcagaattt
     2881 cagataaaaa aaatccttag ctttcgctaa ggatctgaag tggaattcat ga
//
