LOCUS       sequence-9909-ex        3862 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 3862)
  AUTHORS   Zeiser E, Frokjaer-Jensen C, Jorgensen E, Ahringer J
  TITLE     MosSCI and gateway compatible plasmid toolkit for constitutive and 
            inducible expression of transgenes in the C. elegans germline.
  JOURNAL   PLoS One. 2011;6(5):e20082. Epub 2011 May 26.
  PUBMED    21637852
REFERENCE   2  (bases 1 to 3862)
  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: "PLoS One.";
            date: "2011"; volume: "6(5)"; pages: "e20082. Epub 2011 May 26"
COMMENT     Sequence Label: pJA257
FEATURES             Location/Qualifiers
     source          1..3862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(798..826)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1574..1594
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1643..1966
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(1971..2066)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2103..2122)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2104..2122)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2127..2143)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2127..2143)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2245..2264)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2256..3065
                     /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(2323..2342)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3212..3800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3701..3720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggagg gaggtggagg tggagctatg agtaaaggag
      781 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
      841 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
      901 aatttatttg cactactgga aaactacctg ttccatgggt aagtttaaac atatatatac
      961 taactaaccc tgattattta aattttcagc caacacttgt cactactctc acttatggtg
     1021 ttcaatgctt ctcgagatac ccagatcata tgaaacagca tgactttttc aagagtgcca
     1081 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     1141 cacgtaagtt taaacagttc ggtactaact aaccatacat atttaaattt tcaggtgctg
     1201 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     1261 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     1321 acatcatggc agacaaacaa aagaatggaa tcaaagttgt aagtttaaac atgattttac
     1381 taactaacta atctgattta aattttcaga acttcaaaat tagacacaac attgaagatg
     1441 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     1501 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     1561 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     1621 acgaactata caaatgagat aaatgcaaaa tcctttcaag cattcccttc ttctctatca
     1681 ctcttctttc tttttgtcaa aaaattctct cgctaattta tttgcttttt taatgttatt
     1741 attttatgac tttttatagt cactgaaaag tttgcatctg agtgaagtga atgctatcaa
     1801 aatgtgattc tgtctgatgt actttcacaa tctctcttca attccatttt gaagtgcttt
     1861 aaacccgaaa ggttgagaaa aatgcgagcg ctcaaatatt tgtattgtgt tcgttgagtg
     1921 acccaacaaa aagaggaaac tttattgtgc cgccaagaaa aaagtctcag caactttatt
     1981 atacaaagtt ggcattataa aaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2041 tatcagtcaa aataaaatca ttatttggag ctccatggta gcgttaacgc ggccgcgata
     2101 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
     2161 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
     2221 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     2281 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     2341 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     2401 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     2461 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     2521 ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc caggtattag
     2581 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2641 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2701 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2761 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2821 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2881 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2941 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     3001 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     3061 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     3121 acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     3181 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3241 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3301 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3361 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3421 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3481 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3541 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3601 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3661 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3721 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3781 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3841 gctggccttt tgctcacatg tt
//
