LOCUS       sequence-136596-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Saptrap tagRFP + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  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 5483)
  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: pMLS286
FEATURES             Location/Qualifiers
     source          1..5483
                     /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"
     primer_bind     563..582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        563..581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     606..622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     607..623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1208..1241
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3205..3216
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3365..3398
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3656..3672)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3672)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3709..3729)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3709..3727)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3748..3764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3748..3764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3866..3885)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3877..4686
                     /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(3944..3963)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4833..5421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5322..5341
                     /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 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      601 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgctcttcg
      661 atgatggttg tgtcgaaagg cgaagaactg atcaaggaaa acatgcatat gaaactgtac
      721 atggaaggaa ccgtcaataa ccatcacttt aaatgtactt ccgaaggaga aggtaaacca
      781 tacgaaggca cccagactat gcgcattaaa gtcgttgaag gaggtccact gccgtttgcg
      841 ttcgatatcc tggccacctc atttatgtac ggatctagaa ccttcatcaa ccacactcag
      901 ggtatcccag atttctttaa acaatcattt ccggaaggat tcacatggga acgtgtgacc
      961 acttatgaag atggcggagt cctgacagca acgcaggata cgagcctgca agatggttgc
     1021 ctgatttaca atgtcaaaat ccgcggtgtt aattttccaa gtaacggccc ggttatgcag
     1081 aagaaaaccc tgggttggga agcaaacacg gaaatgctgt atccagctga tggtggcctg
     1141 gaaggcagaa ctgatatggc actgaaactg gtgggaggtg ggtaagttta aacatgattt
     1201 tactaacata acttcgtata atgtatgcta tacgaagtta tttctagaca ttctctaatg
     1261 aaaaaatctt tcagttgaaa ttgaaaatga gttaaagttg gagtttttat tgaaaacaga
     1321 tttccgtgtg attagtgttt ttagcgagtg tgacaggaca gcgaaaaaat atagaaacaa
     1381 ggggggaact gaaaagctta ggaatgcatt gaacatgaga aggggaaggg gaaggaacaa
     1441 actagacagg aattattgga atttaatcac atttggagtt ttttttctat tcgacagaat
     1501 aattatccag aacatttttg tattaaatat ttatgcatca tatgagtagt cggctttgtt
     1561 gtgcatgacg agtttgttat cgacgaaata gaagctgtca gaacgagtct cgtttggatt
     1621 gttgatcatg tcgtccactg aaaaagagat tagtcttttg aattgtactt tttagaataa
     1681 tgactcactg agttgttgag agagttgagg gaactcatag atatgttcac agttgtttcg
     1741 tgaattcgga atacagaatc cgaattcaaa gtcaaaacac ttcagaaggc gatctttgaa
     1801 gaagtgacgt tcgatcattc ggaaatgatg gattgggatg tctcctactt tgaattccac
     1861 agttgctccg accgttttga gtttcaaaaa gtttggagca aatctataac gcacgtatct
     1921 tgccgactcc tgtggcgact catcattctc ctgatcgttc tccggtttgg cgatctcgaa
     1981 aagcacttgc tcagtgtcca gatcacggat ttggaacttg gtgaactcga tgttatagat
     2041 gttcgcagat ggggagcata agaatcctaa atttatgttt taaactgaaa tccaaaggga
     2101 gcaagatacc ttgagtgatt cccggaagtg ctaaaacgtc gttcggagtg atttgagctt
     2161 tcttcgcaag ctccgattcc gttgtgattc cttgttcggt gcttggtggt ggccgtggca
     2221 tctggaaata tggaaaagtt caacaaaaag aaaagagaaa agaatgaaat cggatatcaa
     2281 gagttagttg agcggtttct ctagttttct gagtctcacc tgcgacggga aggtcgccga
     2341 gccgggtgga atcgattgtt gttgctcggc tttcatatcg gtttggttgg aagcggctga
     2401 aaacggaaag aagtggaaga aggaaaagag tgtggtgtga caggaaaatg gtaattagag
     2461 ggtgccaaat aaccagctat attttgtttt tttttgaaaa catttttaaa aagaaaaata
     2521 cgataatgat atcagatgga tttccggaaa actggtatga aaaatttcaa cctttttgag
     2581 tacatgtaat caaaatacac tttgtaaatt atcattttta ttgaaactcc accatttttc
     2641 tatttataac gctaataatt tgaaaaagaa acctgttgcg aaccgcgggg tgaatcccaa
     2701 aaacgaatgc gttttggtgg agtgattgat tcgaatcgaa gaagaaaaag aagaagacgt
     2761 ggaatagaga gctcactctt aaccgagcag cacacaccga cagaaaaaaa aatgaaatga
     2821 atgagggtct tcttcttctt cttcttcgaa tgattgacag aaatgggaaa aagaggaaga
     2881 ttgagaaggg aaaaaggaag gagaaaagaa gcagaagaag acgtcagaga ggagaggaac
     2941 gagcggaaaa gcagcgggcg caagtcatag aagtagcaga gctggggaga agaagacact
     3001 atccaagaaa ggaatgacga gagagtatgc aaaggggtat agggtgcaga cagaatagga
     3061 acagaataac agatgatgag ccaagaagag ttgaaaaggg cgatgaattt gtcatgtaac
     3121 ttaatttggg tcaatttgag catgatgaat tgaaatcatc ccttgttggg agttaataac
     3181 cggtttgtta tcagaaaccc tgtaatagaa gggcgcccta actttgagcc aattcatccc
     3241 ggtttctgtc aaatatatca aaaagtggtc aactgacaaa ttgtttttga tattataata
     3301 aacattttat ccgttaacaa ttttcgaata ctttttacaa ggacttggat aaattggctc
     3361 aaagataact tcgtataatg tatgctatac gaagttatta actaatctga tttaaatttt
     3421 cagccatctg atctgtaact tcaagacaac gtatcgttct aaaaaaccag cgaaaaacct
     3481 gaaaatgccg ggagtttatt acgtggatca ccgtctggaa cgcatcaaag aagccgataa
     3541 agaaacctat gtggaacaac atgaagtggc agtcgctcgt tactgcgatc tgccgagcaa
     3601 actgggccac aaactgaatg gaatggatga actgtacaaa ggtcgaagag cgggggatcc
     3661 actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg
     3721 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctggcagct ctggcccgtg
     3781 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     3841 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     3901 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     3961 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     4021 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4081 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4141 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     4201 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4261 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     4321 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4381 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4441 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4501 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4561 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4621 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     4681 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     4741 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     4801 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4861 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4921 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4981 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5041 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5101 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5161 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5221 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5281 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5341 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5401 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5461 tgctggcctt ttgctcacat gtt
//
