LOCUS       Exported                3124 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  SapTrap syntron-embedded FLP-on N-terminal connector donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pMLS281
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3124)
  AUTHORS   Schwartz ML, Jorgensen EM
  TITLE     SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification
            in Caenorhabditis elegans.
  JOURNAL   Genetics. 2016 Feb 2. pii: genetics.115.184275.
  PUBMED    26837755
REFERENCE   2  (bases 1 to 3124)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3124
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    711..744
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1142..1175
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1249..1348)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1365..1384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1366..1384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1389..1405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1389..1405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1507..1526)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1518..2327
                     /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(1585..1604)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2474..3062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2963..2982
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctat gctcttcggg ttcggtaagt ttaaacatga ttttactaac gaagttccta
      721 ttctctagaa agtataggaa cttctaacta atctgattta aattttcagg gtagccatgg
      781 cttcccgccg gaggtggagg agcaggatga tggcacgctg cccatgtctt gtgcccagga
      841 gagcgggatg gaccgtcacc ctgcagcctg tgcttctgct aggatcaatg tgtaagctgt
      901 ctcatcctac tttcacctag ttaactgctt gtcttaaaat ctatgcttct ctttagtatc
      961 taaaattttc ctagaagctt acaagtatat aaatggtctc ttctcaataa aggttgtata
     1021 tttattcatc ttattgaatc tgccatttcc tcgtttttgc gagtttatat accttccaat
     1081 tttctttcta ttgtattttc aacttctaat tttaattcag ggaaactgct tcaacgcatc
     1141 agaagttcct attctctaga aagtatagga acttctaact aatctgattt aaattttcag
     1201 ggtatgggca gcggaggtac cggcggtagt ggaggcacgc gaagagctac ccagctttct
     1261 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1321 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     1381 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     1441 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     1501 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     1561 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1621 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1681 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1741 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1801 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag
     1861 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1921 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1981 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2041 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     2101 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2161 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2221 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2281 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     2341 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     2401 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     2461 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2521 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2581 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2641 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2701 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2761 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2821 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2881 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2941 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3001 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3061 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3121 tgtt
//
