LOCUS       sequence-309411-        3500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA Destination Vector for SapTrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3500)
  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 3500)
  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: pJW1838
FEATURES             Location/Qualifiers
     source          1..3500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..57)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     157..176
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        408..907
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_feature    1028..1264
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1283..1299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1283..1299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1296..1318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1307..1323
                     /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(1331..1337,1338..1355,1356..1361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1376..1397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1516..1533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1687..2275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1767..1786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2446..3237,3238..3306))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3069..3088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3307..3411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3479..3497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
       61 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      121 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      181 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      241 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcctc caagaactcg
      421 tacaaaaatg ctctgaagta ggtctcgaga tcaatactgg gaagacgaaa gtcttgcgaa
      481 accgattcgc tgaccccagt aaagtctact tcggtagccc ttcccccacc acccagctcg
      541 acgacgtcga cgagtacatc tacctcggtc gtcaaatcaa cgcccaaaac aacttgatgc
      601 cggaaatcca ccgaagacgt cgagcagcct gggctgcatt caatggaatc aagaatgcca
      661 ccgactccat caccgacaag aagattcgtg cgaatctgtt cgactcaatt gtccttccag
      721 cgctcaccta cgtttcagaa gcctggacat tcaccaaagc tctatccgaa cgagtacgaa
      781 tcacacatgc ctcccccaca aattattgat ctacatcccc atttggtata gtgtcttaca
      841 ggactctcaa tatgtcagct gcatgataaa tggtccctat ataattgaat tgcaaatcta
      901 aatgtttgcg aagagcggat ccatgggctc ttcggtttaa gagctatgct ggaaacagca
      961 tagcaagttt aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     1021 tttttttcaa taatattcta gatatccctt tttgaaacta aaatgaaata ctctaaattt
     1081 ttgagttcaa tgtttcaact aatcttggtt aaaggtaaat tcccggaggg acgattttaa
     1141 agttgccaga gttgattaaa ttggtccaaa aaattttatt aggaaactcg aaaaaaaatt
     1201 gatttttttt gacaatattc caaagaaaat tacaaaatct cgacgccaaa catagtcggc
     1261 tgtgaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1321 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1381 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1441 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1501 gctcatatgc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2101 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2341 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2401 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2461 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2521 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2581 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2641 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2701 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2761 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2821 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2881 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2941 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3001 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3061 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3121 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3181 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3241 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3301 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3361 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3421 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3481 taggcgtatc acgaggccct
//
