LOCUS       sequence-309434-        3842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NT Donor-cassette for SapTrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3842)
  AUTHORS   Ashley GE, Duong T, Levenson MT, Martinez MAQ, Johnson LC, Hibshman 
            JD, Saeger HN, Palmisano NJ, Doonan R, Martinez-Mendez R, Davidson 
            BR, 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 Mar 31;217(3). pii: 6104563. doi: 
            10.1093/genetics/iyab006.
  PUBMED    33677541
REFERENCE   2  (bases 1 to 3842)
  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.";
            date: "2021-03-31"; volume: "217(3). pii: 6104563. doi"; pages: " 
            10.1093/genetics/iyab006"
COMMENT     Sequence Label: pJW1759
FEATURES             Location/Qualifiers
     source          1..3842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(98..116)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(99..116)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(134..150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(134..150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(147..169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    158..174
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(182..188,189..206,207..212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    227..248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(365..382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(536..1124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(616..635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1262..1636)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      1633..1642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1842..2636)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1842..2636)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1953..1972)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2563..2582
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2985..3287)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(3087..3106)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3410..3432
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3424..3441
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3425..3441
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3448..3467
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3448..3466
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3574..3594
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3616..3747
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     CDS             3775..3840
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 gcgaagagcg acccagctaa gggcgaattc cagcacactg gcggccgtta ctagtggatc
       61 cgagctcggt accaagcttg atgcatagct tgagtattct atagtgtcac ctaaatagct
      121 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      181 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
      241 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      301 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      361 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      421 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      481 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      541 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      601 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      661 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      721 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      781 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      841 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      901 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      961 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1021 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1081 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1141 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1201 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttagcacgt
     1261 gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga
     1321 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga
     1381 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca
     1441 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
     1501 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
     1561 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
     1621 tggtcaactt ggccatggtg gccctcctca cgtgctatta ttgaagcatt tatcagggtt
     1681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1741 cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta
     1801 aggagaaaat accgcatcag gaaattgtaa gcgttaataa ttcagaagaa ctcgtcaaga
     1861 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     1921 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     1981 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     2041 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     2101 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     2161 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     2221 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     2281 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     2341 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     2401 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     2461 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     2521 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     2581 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     2641 aacgatcctc atcctgtctc ttgatcagag cttgatcccc tgcgccatca gatccttggc
     2701 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     2761 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     2821 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     2881 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgaaa
     2941 aggatctagg tgaagatcct ttttgataat ctcatgcctg acatttatat tccccagaac
     3001 atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc
     3061 ccgataacgg agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc
     3121 ccgatatgca ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg
     3181 gccaggggga tcaccatccg tcgccccggc gtgtcaataa tatcactctg tacatccaca
     3241 aacagacgat aacggctctc tcttttatag gtgtaaacct taaactgccg tacgtatagg
     3301 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3361 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3421 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attgggccct
     3481 ctagatgcat gctcgagcgg ccgccagtgt gatggatatc tgcagaattc gcccttgctg
     3541 ggtcctagct cttcgggtgg cggcagcgga ggagagaatc tgtactttca atccggatcc
     3601 ggaggtggcg ggatgcctaa agatccagcc aaacctccgg ccaaggcaca agttgtggga
     3661 tggccaccgg tgagatcata ccggaagaac gtgatggttt cctgccaaaa atcaagcggt
     3721 ggcccggagg cggcggcgtt cgtgaagggt ggtggcggtt ctggctcagg atctgactac
     3781 aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga cgatgacaag
     3841 ac
//
