LOCUS       sequence-12199-e        2927 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 2927)
  TITLE     J.C. Anderson Lab Plasmids
REFERENCE   2  (bases 1 to 2927)
  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-J72020
FEATURES             Location/Qualifiers
     source          1..2927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             45..71
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(101..118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    complement(180..204)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        251..355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(356..424,425..1216)
                     /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(574..593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(1497..1516)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1508..2323
                     /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             1508..2323
                     /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(1581..1600)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2406..2762
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2824..2848
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gatcttaagc acccatagaa tacacacctg gaggaataat gtcttaccca tacgacgtcc
       61 cagactacgc tgggggatcc taactcgctc ctcaggcttc ctcgctcact gactcgctgc
      121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atcaattcga
      181 cccagctttc ttgtacaaag tggttgatcg gatctcttag acgtcaggtg gcacttttcg
      241 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      301 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      361 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      421 tgctcaccca gaaacgctgg tgaaagtaaa agatgcagag gatcagttgg gtgcacgagt
      481 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      541 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      601 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      661 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      721 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      781 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      841 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
      901 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      961 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1021 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1081 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1141 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1201 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1261 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc ggatctctga
     1321 agactctcga gctgcaggac tcacagctcc tcggatctgc tgatccttca actcagcaaa
     1381 agttcgattt attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca
     1441 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1501 gggtgttatg agccatattc aacgggaaac gtcttgctcc aggccgcgat taaattccaa
     1561 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1621 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1681 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     1741 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1801 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1861 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1921 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1981 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2041 ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2101 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2161 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2221 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2281 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     2341 gttgtaacac tggcagagca ttacgctgac ttgacgggag gatctagcag ttcaacctgt
     2401 tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa
     2461 gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg
     2521 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa
     2581 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa
     2641 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact
     2701 gagaagccct tagagcctct caaagcaatt ttgagtgaca caggaacact taacggctga
     2761 catgggaatt agccatgggc ccgtgcgaat cacggatctc ccgatcaaga attgccatta
     2821 tcaacaagtt tgtacaaaaa agcaggctcc gaattggtat cacgaggcag aatttcagat
     2881 aaaaaaaatc cttagctttc gctaaggatc tgaagtggaa ttcatga
//
