LOCUS       sequence-309448-        3685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor-cassette for SapTrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3685)
  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 3685)
  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: pJW1347
FEATURES             Location/Qualifiers
     source          1..3685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    72..93
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(108..113,114..131,132..138)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    146..162
                     /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)."
     primer_bind     151..173
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     170..186
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     170..186
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        204..222
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     204..221
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(538..557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(539..557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(564..581)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(564..580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(573..595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             718..1020
                     /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     899..918
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1369..2163
                     /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             1369..2163
                     /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(1423..1442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2033..2052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2363..2372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2369..2743
                     /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"
     rep_origin      2881..3469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3370..3389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3623..3640
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
       61 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
      121 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
      181 tatgaccatg attacgccaa gctatttagg tgacactata gaatactcaa gctatgcatc
      241 aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcgccct
      301 tctttcctgc gttatcccct gattctgtgg ctcttcggcg ggtggcggtg gatcgggagg
      361 aggaggttcg ggtggcggag gcagtggagg tggcggaagt ggcggcggtg gttcaggagg
      421 aggcggatcc atgcgaagag cgtttgatgc ctggcagttt atggcgggaa gggcgaattc
      481 tgcagatatc catcacactg gcggccgctc gagcatgcat ctagagggcc caattcgccc
      541 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      601 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      661 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctata cgtacggcag
      721 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
      781 gatattattg acacgccggg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
      841 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
      901 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
      961 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     1021 atgtcaggca tgagattatc aaaaaggatc ttcacctaga tccttttcac gtagaaagcc
     1081 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     1141 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     1201 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     1261 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     1321 cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     1381 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     1441 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     1501 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     1561 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     1621 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     1681 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1741 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     1801 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     1861 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     1921 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1981 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2041 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2101 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2161 tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     2221 atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2341 tcaataatag cacgtgagga gggccaccat ggccaagttg accagtgccg ttccggtgct
     2401 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     2461 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     2521 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     2581 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     2641 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     2701 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc taaaacttca
     2761 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3541 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3601 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3661 acgcaaaccg cctctccccg cgcgt
//
