LOCUS       sequence-8546-ex        3164 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 3164)
  AUTHORS   Merritt C, Seydoux G
  TITLE     Transgenic solutions for the germline.
  JOURNAL   WormBook. 2010 Feb 8:1-21. doi: 10.1895/wormbook.1.148.1.
  PUBMED    20169625
REFERENCE   2  (bases 1 to 3164)
  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: "WormBook. 
            2010 Feb 8:1-21. doi: 10.1895/wormbook.1.148.1."
COMMENT     Sequence Label: pCM1.53
FEATURES             Location/Qualifiers
     source          1..3164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(73..100)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(192..278)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    331..430
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(463..491)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1239..1259
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(1340..1439)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1551..1570)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1562..2371
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1629..1648)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2514..3102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3003..3022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagcc aggaagagtt
       61 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      121 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      181 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      241 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      301 cgcgttaacg ctagcatgga tctcgggccc caaataatga ttttattttg actgatagtg
      361 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaag tttgtacaaa
      421 aaagcaggct caatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
      481 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
      541 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
      601 tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca
      661 cttgtcacta ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa
      721 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
      781 ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca
      841 tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga
      901 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
      961 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     1021 gttgtaagtt taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc
     1081 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat
     1141 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     1201 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     1261 gctgctggga ttacacatgg catggacgaa ctatacaaaa gcccacaagc tttggaattc
     1321 gctagccggc catacaagaa cccagctttc ttgtacaaag tgggcattat aagaaagcat
     1381 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1441 catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1501 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1561 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1621 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1681 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1741 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1801 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1861 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1921 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1981 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2041 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2101 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2161 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2221 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2281 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2341 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2401 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2461 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2521 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2581 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2641 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2701 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2761 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2821 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2881 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2941 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3001 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3061 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
