LOCUS       sequence-272307-        3341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C. elegans KNL1 with 8A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3341)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3341)
  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            "Unpublished"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: pDK324
FEATURES             Location/Qualifiers
     source          1..3341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     230..247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     332..351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        332..350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             382..404
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             424..441
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1054..1072)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1068..1115
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1212..1667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1509..1530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1693..1797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1798..1866,1867..2658)
                     /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(2016..2035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2829..3341,1..76)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3318..3337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
       61 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      181 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      241 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      301 tcattaatgc aggatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
      361 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt
      421 tctcatcatc atcatcatca tggtatggct agcactgaca ctattggatc agatggtatt
      481 ttgacaccac gtggcggaaa catggatatt tccgaatctc cggcctgcac gtcctcattt
      541 caagtgttcg gcggtggtaa tctcgataaa actatggata tgtctctcga aacaactatc
      601 aacgagaaca acgaaacggc gagattgttt gaaaccacaa gagatccaac agctgctgca
      661 gagaaagctg ccgaaaccac tactaaggct actgaacgtg ctgctagtat gccactggat
      721 gataccttag caatgttcaa tacaacgaat caagaagata aggatatgtc agttgatcgt
      781 tcagttcttt tcacgattcc caaagttccg aagcataacg ctacaatgaa tagaactata
      841 ccgatggacc tcgatgaatc aaaagcagcg ggcggccagt gcgatgaaac gatgaatgtg
      901 ttcaatttca caaacttgga agccgctgaa atggatacga gtaaattaga tgaataggga
      961 tccgagctcg agatctgcag ctggtaccat ggaattcgaa gcttgatccg gctgctaaca
     1021 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     1081 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     1141 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1201 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1261 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1321 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1381 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1441 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1501 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1561 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1621 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     1681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2041 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2341 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2941 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3301 acgagggagc ttccaggggg aaacgcctgg tatctttata g
//
