LOCUS       sequence-136622-        2603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap TEV-Flagtag N-terminal connector donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2603)
  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 2603)
  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: "Genetics. 
            2016 Feb 2. pii: genetics.115.184275."
COMMENT     Sequence Label: pMLS383
FEATURES             Location/Qualifiers
     source          1..2603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     109..125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     144..164
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        146..164
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     201..217
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     201..217
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             253..273
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             286..309
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(370..386)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(371..387)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(411..430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(412..430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(440..457)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(440..456)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(449..471)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      520..606
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      698..725
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(884..901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1055..1643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1135..1154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1790..2599)
                     /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     2513..2532
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     join(2591..2603,1..7)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
       61 tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcca ggaaacagct
      121 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
      181 tccaccgcgg tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgatc
      241 tgctcttcgg gtgagaatct gtactttcag agcgagccaa ccactgacta caaggacgac
      301 gacgacaaga gtactagcgg tggcagtgga ggtaccggcg gaagcacggg aagagcacat
      361 caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg ccctatagtg
      421 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc
      481 atgctagcgt taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
      541 tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
      601 acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca
      661 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc
      721 ctttttgcgt ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct
      781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1681 agtggaacga cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc
     1741 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     1801 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     1861 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     1921 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     1981 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2041 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2101 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     2161 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     2221 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     2281 cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     2341 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     2401 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     2461 catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     2521 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt
     2581 cccgttgaat atggctcata aca
//
