LOCUS       Exported                9225 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Multi-cassette for SapTrap.
ACCESSION   .
VERSION     .
KEYWORDS    pJW1846
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9225)
  AUTHORS   Ashley G, Duong T, Levenson MT, Martinez MAQ, Johnsen LC, Hibshman 
            JD, Saeger HN, Palmisano NJ, Doonan R, Martinez-Mendez R, Davidson 
            B, Zhang W, Ragle JM, Medwig-Kinney TN, Sirota SS, Goldstein B, 
            Matus DQ, Dickinson DJ, Reiner DJ, Ward JD
  TITLE     An expanded auxin-inducible degron toolkit for Caenorhabditis 
            elegans
  JOURNAL   Genetics, 2021
REFERENCE   2  (bases 1 to 9225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9225
                     /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    1553..1586
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             1596..1625
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3442..3765
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    5683..5707
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5710..6735
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(6736..6760)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    7113..7146
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             7171..7191
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             7372..7437
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7466..7485)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7467..7485)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7490..7506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7490..7506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7608..7627)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7619..8428
                     /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(7686..7705)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8575..9163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9064..9083
                     /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 aataatgctc ttcggcgggt ggcggtggat
      601 cgggaggagg aggttcgggt ggcggaggca gtggaggtgg cggaagtggc ggcggtggtt
      661 caggaggagg cggatccatg agtaaaggag aagaattgtt cactggagtt gtcccaatcc
      721 tcgtcgagct cgacggagac gtcaacggac acaagttctc cgtctccgga gagggagagg
      781 gagacgccac ctacggaaag ctcaccctca agttcatctg caccaccgga aagctcccag
      841 tcccatggcc aaccctcgtc accaccttct gctacggagt ccaatgcttc tcccgttacc
      901 cagaccacat gaagcgtcac gacttcttca agtccgccat gccagaggga tacgtccaag
      961 agcgtaccat cttcttcaag gtaagtttaa acatatatat actaactact gattatttaa
     1021 attttcagga cgacggaaac tacaagaccc gtgccgaggt caagttcgag ggagacaccc
     1081 tcgtcaaccg tatcgagctc aaggtaagtt taaacagttc ggtactaact aaccatacat
     1141 atttaaattt tcagggaatc gacttcaagg aggacggaaa catcctcgga cacaagctcg
     1201 agtacaacta caactcccac aacgtctaca tcatggccga caagcaaaag aacggaatca
     1261 aggtcaactt caaggtaagt ttaaacatga ttttactaac taactaatct gatttaaatt
     1321 ttcagatccg tcacaacatc gaggacggat ccgtccaact cgccgaccac taccaacaaa
     1381 acaccccaat cggagacgga ccagtcctcc tcccagacaa ccactacctc tccacccaat
     1441 ccgccctctc caaggaccca aacgagaagc gtgaccacat ggtcctcctc gagttcgtca
     1501 ccgccgccgg aatcacccac ggaatggacg agctctacaa ggtaagttta aaataacttc
     1561 gtatagcata cattatacga agttattttc agggagagca aaagcttatc tccgaggagg
     1621 acctctaagg atgatcgacg ccaacgtcgt tgaattttca aattttaaat actgaatatt
     1681 tgtttttttt cctattattt atttattctc tttgtgtttt ttttcttgct ttctaaaaaa
     1741 ttaattcaat ccaaatctaa acattttttt ttctctttcc gtctcccaat tcgtattccg
     1801 ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg
     1861 acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg
     1921 acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat
     1981 aaaaatatat agaggaggaa ggaaactagc tgccctcgat tgtctatctt ccattctcac
     2041 caagtttcaa caactttatt cagtcattct tctctggttc atttcccaac caatcttccc
     2101 gaataaagcc catctgtcct ttcggccaca ttcctcacct acatccgtac tccttatctc
     2161 ccgttcttat ccagtctttt atcgtctctg cgtctcgcac actgtctcgt cggtaatgta
     2221 aaaaaaaaac aaaaaaaact tgaaaacttg cggaaagatt tcctaataaa aggagacgaa
     2281 tccggtagga aatcgacaag tctctgcttc tcgaagacta ggtgagttac atttctttgg
     2341 ccgaattatt ttgaaatcct aaatttgata atttcagaga tgtctgtaaa acttgcgtgt
     2401 tatgtgacgg cttcggtcac tgtcgccact cttatggtat gcttcatgac catgtcaacc
     2461 atctactcgg aggttgatgg attcagagag aagcttgata ccgagatgaa tgtgttcaga
     2521 gtgagttaaa ttatattttg aattttaaat aatttttaat tttctagcaa tctaccaatg
     2581 gattgtggaa ggacatagtt gtcatcggaa gatctagcaa gcgtgtccgt tgtcaatatg
     2641 aagagaccaa cgctacccca actccacatg ctgatggatc cccatctgct ccaccaggtc
     2701 aaccaccagc agttccacca gtcttcaacc agccaaagac tccaaatgga gccaatggaa
     2761 atggaccaac ctgcaactgc aatgctgata acaagtgccc agctggacca tccggaccaa
     2821 agggagttcc aggagttcca ggactcgacg gagttccagg acttgacggt gttccaggag
     2881 ttggagctga tgatatcgct ccacaacgcg agtctgtcgg atgcttcact tgcccacaag
     2941 gaccagttgg accaccagga gctcttggaa gaccaggacc acgtggactt ccaggaccaa
     3001 gaggacaaaa tggaaaccca ggaagagatg gacaaccagg acatccagga gagcaaggat
     3061 catccggtca aatcggaaag atcggagagc caggaccacc aggagagaag ggacgcgacg
     3121 ccgagcatcc aatcggaaga ccaggaccaa agggaccaag aggagatcaa ggaccaacag
     3181 gaccagctgg acagaacggt cttcacggac caccaggaga gccaggaacc gttggaccag
     3241 aaggaccatc tggaaagcaa ggacgtcaag gaccagacgg aacccaggga gagactggac
     3301 cagacggaag accaggaaag gatgccgagt actgccagtg cccagacaag tctccaccat
     3361 cagaggctgt caacgccaac cgtggataca gaaatatcta aattgttggt gttttctaat
     3421 aaaatatttg agattcagtg agactttttt cttggcggca caataaagtt tcctcttttt
     3481 gttgggtcac tcaacgaaca caatacaaat atttgagcgc tcgcattttt ctcaaccttt
     3541 cgggtttaaa gcacttcaaa atggaattga agagagattg tgaaagtaca tcagacagaa
     3601 tcacattttg atagcattca cttcactcag atgcaaactt ttcagtgact ataaaaagtc
     3661 ataaaataat aacattaaaa aagcaaataa attagcgaga gaattttttg acaaaaagaa
     3721 agaagagtga tagagaagaa gggaatgctt gaaaggattt tgcatttatc ttaatcaccg
     3781 tcttcgagaa gacgaaccat ggcgccagtt tcagaatcca agtttctaat atagttcatc
     3841 acaatgttga cgttggtcca acctccggct tgcataatct ctgggatgga aactccagcg
     3901 cgagccatgt cacgggcagc tccgacacgg gcggaatgtc cagaccaagc caagtaacgt
     3961 tgtccggaat catccttggc tccgtagatg agacggtggg tagcttcgaa gattcctgaa
     4021 aatttatttt cagcttattc ttagctaata tgttgtttaa acttaccttc aagagctcga
     4081 gtcgagagtt gtgatgttgc actcggggca gcaactccgt tcttgcgaac acgacagaac
     4141 aggtagttgt ttggatcatc ggcgactccg gaaaccgaaa tccagcgctc aacgagcttg
     4201 gtaactccaa gggagagcgc tttttcgact ccggcagtcg agaccagcgt ttttgtacgt
     4261 ccgatgtgga tgagcattct tcctccgtct gtgcgactaa tatccttgac gcgaatgcgg
     4321 gcaatttcgg cgatacggag aagtgtgttg taagcaattc cgaggaaggc gaggtttcgg
     4381 atgtcctggc aacgatcgga attttccata agagaacgga cctgatcgaa gtcggttctg
     4441 aaaatttcgt caagtcttag gattgaatac attaatattt aaacttacct ttcgaaggcg
     4501 agggcttgct tggctctctc tccagcatca acattttcct tgcggatacg tctcattacg
     4561 agagatacgg cattagaatc tgatgggcgt ggaagtccag atcggcgatg gagcatattg
     4621 agctgtccga gatgttgttg gatcgtcttc accgccaatc cacgagcttg gaggtagagc
     4681 aagtagtcac gaacgtcttc tggctcggct gggaaccact tgcggttgtt aagcttgcac
     4741 caggcagccc aagaacggca cactgagaga agcatcttcc aggtgtgctc ggagaaggct
     4801 tgtctgtcac ggaacatatc catgaggttc ttgcgaactt catcacttgt cgcgtcaact
     4861 ggaagtgctg gaagattttg atgaacagtg agaagatttg acatattttc gaagtttttt
     4921 agatgcacta gaacaaagcg tgttggcttc ctctgagccc gctttcctta tatacccgca
     4981 ttctgcagcc ttacagaatg ttctagaagg tcctagatgc attcgtttga aaatactccc
     5041 ggtgggtgca aagagacgca gacggaaaat gtatctgggt ctctttattg tgtacactac
     5101 ttttccatgt accgaatgtg agtcgccctc cttttgcaac aagcagctcg aatgttctag
     5161 aaaaaggtgg aaaatagtat aaataccgtt gaaaataaat accgaacaac atttgctcta
     5221 attgtgaaat tagaaatctt caaactataa tcatgtcact ttaccactat ttccgtccag
     5281 ctcaacgttc cgtttttggt gatcattttt gctttcgtcg taaatctaca cacgcgtctc
     5341 ttccgtgcga gagtccaagc cagcagccaa attcgttgac tgagtattca acgtttatac
     5401 gttgtcggca acgagaaata ggaaaatgca tcgggaaatg ttcttttttc gattttttcc
     5461 aaggttttga caaattttac cacgaatttt gctatgtttt caattaaaaa atatgttatt
     5521 caactgtttc tatgaggaaa ataaggcttt gcatgtaatt ttcttattca gcataatttt
     5581 taattaattt gaattttctg tcctaacgtt tattttgttt tcttggttat gactgatctg
     5641 aaattaattt ttgaatttta aggtaatatg tcaggcggtg ccgcaagttt gtacaaaaaa
     5701 gcaggctcca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     5761 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     5821 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     5881 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     5941 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     6001 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     6061 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     6121 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     6181 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     6241 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     6301 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     6361 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     6421 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     6481 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     6541 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     6601 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     6661 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     6721 agggcaaagg aatagaccca gctttcttgt acaaagtggg tccaattact cttcaacatc
     6781 cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc
     6841 tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga
     6901 ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat
     6961 taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta
     7021 cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata
     7081 tgttacgttt cagtttatga ccgcaatttt taataacttc gtatagcata cattatacga
     7141 agttattttc agggtggcgg cagcggagga gagaatctgt actttcaatc cggatccgga
     7201 ggtggcggga tgcctaaaga tccagccaaa cctccggcca aggcacaagt tgtgggatgg
     7261 ccaccggtga gatcataccg gaagaacgtg atggtttcct gccaaaaatc aagcggtggc
     7321 ccggaggcgg cggcgttcgt gaagggtggt ggcggttctg gctcaggatc tgactacaaa
     7381 gaccatgacg gtgattataa agatcatgat atcgattaca aggatgacga tgacaagacg
     7441 cgaagagcgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     7501 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     7561 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     7621 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     7681 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     7741 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     7801 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     7861 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     7921 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     7981 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     8041 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     8101 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     8161 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     8221 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     8281 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     8341 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     8401 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     8461 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     8521 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
