LOCUS       sequence-251926-        3558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant of pKD13. Lambda Red recombineering PCR template plasmid 
            carrying an F5-tetA-F5 cassette orthogonal to FRT-KanR-FRT in pKD13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3558)
  TITLE     Lambda Red recombineering PCR template plasmids, variants of pKD13 
            with orthogonal marker-FRT sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3558)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pES133
FEATURES             Location/Qualifiers
     source          1..3558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..247)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     promoter        join(372..377,378..394,395..400)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             448..1638
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(800..819)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      complement(1748..1775)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1867..1953)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2043..2147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2148..2216,2217..3008)
                     /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(2366..2385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3110..3498
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctactggtat tggcacaaac ctgattccaa tttgagcaag gctatgtgcc atctcgatac
       61 tcgttcttaa ctcaacagaa gatgctttgt gcatacagcc cctcgtttat tatttatctc
      121 ctcagccagc cgctgtgctt tcagtggatt tcggataaca gaaaggccgg gaaataccca
      181 gcctcgcttt gtaacggagt agagacgaaa gtgattgcgc ctacccggat attatcgtga
      241 ggatgcgtca tcgccattaa ttcactgatc agtgataagc tgtcaaacat gagaattaat
      301 tccggggatc cgtcgacctg cagttcgaag ttcctattct tcaaaaggta taggaacttc
      361 aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta
      421 aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc
      481 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
      541 ttgcgggaca tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta
      601 tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc
      661 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg
      721 accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat caccggcgcc
      781 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
      841 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
      901 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
      961 ctcaacctac tactgggctg ctttctaatg caggagtcgc ataagggaga gcgtcgaccg
     1021 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     1081 gtcgccgcac ttatgactgt attctttatc atgcaactcg taggacaggt gccggcagcg
     1141 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     1201 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     1261 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     1321 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     1381 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     1441 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga
     1501 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     1561 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     1621 cgggccacct cgacctgaag ttcctattct tcaaaaggta taggaacttc gaagcagctc
     1681 cagcctacac aatcgctcaa gacgtgtaat gctgcaatct gcatgcaagc ttggcactgg
     1741 ccacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
     1801 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
     1861 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     1921 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
     1981 ggggagaccc cacactacca tcgggggcca tcgatgcagg tggcactttt cggggaaatg
     2041 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2101 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2161 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2221 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2281 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2341 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2401 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2461 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2521 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2581 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2641 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2701 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2761 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2821 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2881 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2941 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3001 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3061 tttaattttt gcggccgcaa gatccggcca cgatgcgtcc ggcgtagagg atctgaagat
     3121 cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc
     3181 atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa
     3241 gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac
     3301 aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa
     3361 aaaaaagaat atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc
     3421 agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg
     3481 aacacttaac ggctgacatg ggaattagcc atggcaaagc ctcgcaatcc agtgcaaagc
     3541 tagcttcttc gtctgttt
//
