LOCUS       sequence-370753-        4009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template for repair templates to tag a gene with a mKate2 and 
            mIAA7 AID degron using homology-directed repair for C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4009)
  AUTHORS   Sepers JJ, Verstappen NHM, Vo AA, Ragle JM, Ruijtenberg S, Ward JD, 
            Boxem M
  TITLE     The mIAA7 degron improves auxin-mediated degradation in C. elegans.
  JOURNAL   G3 (Bethesda). 2022 Aug 27. pii: 6677660. doi: 
            10.1093/g3journal/jkac222.
  PUBMED    36029236
REFERENCE   2  (bases 1 to 4009)
  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: "G3 
            (Bethesda). 2022 Aug 27. pii: 6677660. doi: 
            10.1093/g3journal/jkac222."
COMMENT     Sequence Label: pJJS009
FEATURES             Location/Qualifiers
     source          1..4009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1824..1842)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1863..1879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1863..1879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1876..1898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1887..1903
                     /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(1911..1917,1918..1935,1936..1941))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1956..1977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2094..2111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3024..3815,3816..3884))
                     /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     3647..3666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3885..3989)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggatgggat
      661 tctccgagac cgtcgacctc atgctcaacc tccaatccaa caaggaggga tccgtcgacc
      721 tcaagaacgt ctccgccgtc ccaaaggaga agaccaccct caaggtaagt ttaaacatat
      781 atatactaac taaccctgat tatttaaatt ttcaggaccc atccaagcca ccagccaagg
      841 cccaagtcgt cggatggcca ccagtccgta actaccgtaa gaacatgatg acccaacaaa
      901 agacctcctc catggtctcc gagctcatta aagaaaacat gcatatgaag ctctacatgg
      961 agggaaccgt caacaaccac cacttcaagt gcacctccga gggagaggga aagccatacg
     1021 agggaaccca aaccatgcgt atcaaggccg tcgagggagg accactccca ttcgccttcg
     1081 acatcctcgc cacctccttc atgtacggat ccaaggtaag tttaaacata tatatactaa
     1141 ctaaccctga ttatttaaat tttcagacct tcatcaacca cacccaagga atcccagact
     1201 tcttcaagca atccttccca gagggattca cctgggagcg tgtcaccacc tacgaggacg
     1261 gaggagtcct caccgccacc caagacacct ccctccaaga cggatgcctc atctacaacg
     1321 tcaaggtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcagatcc
     1381 gtggagtcaa cttcccatcc aacggaccag tcatgcaaaa gaagaccctc ggatgggagg
     1441 cctccaccga gaccctctac ccagccgacg gaggacttga gggacgtgcc gacatggccc
     1501 tcaagctcgt cggaggagga cacctcatct gcaacctcaa ggtaagttta aacatgattt
     1561 tactaactaa ctaatctgat ttaaattttc agaccaccta ccgttccaag aagccagcca
     1621 agaacctcaa gatgccagga gtctactacg tcgaccgtcg tcttgagcgt atcaaggagg
     1681 ccgacaagga gacctacgtc gagcaacacg aggtcgccgt cgcccgttac tgcgacctcc
     1741 catccaagct cggacaccgt aagggcagcg gtggcagtgg aggtaccggc ggaagcggtg
     1801 cggtggagct ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca
     1861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgc
//
