LOCUS       sequence-370750-        4012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template for repair templates to tag a gene with a mCherry 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 4012)
  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 4012)
  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: pJJS004
FEATURES             Location/Qualifiers
     source          1..4012
                     /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(1827..1845)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1866..1882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1879..1901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1890..1906
                     /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(1914..1920,1921..1938,1939..1944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1959..1980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2097..2114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2268..2856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2348..2367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3027..3818,3819..3887))
                     /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     3650..3669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3888..3992)
                     /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 tggggcagcg
      661 gtggcagtgg aggtaccggc ggaagcggta tgtccaaggg agaggaggac aacatggcca
      721 tcatcaagga gttcatgcgt ttcaaggtcc acatggaggg atcagtcaac ggacacgagt
      781 tcgagatcga gggagaggga gagggacgtc catacgaggg aacccaaacc gccaagctca
      841 aggtaagttt aaacatatat atactaacta accctgatta tttaaatttt caggtcacca
      901 agggaggacc actcccattc gcctgggaca tcctctcccc acaattcatg tacggatcaa
      961 aggcctacgt caagcaccca gccgacatcc cagactacct caagctctcc ttcccagagg
     1021 gattcaagtg ggagcgtgtc atgaacttcg aggacggagg agtcgtcacc gtcacccaag
     1081 actcctccct ccaagacgga gagttcatct acaaggtaag tttaaacagt tcggtactaa
     1141 ctaaccatac atatttaaat tttcaggtca agctccgtgg aaccaacttc ccatccgacg
     1201 gaccagtcat gcaaaagaag accatgggat gggaggcctc ctccgagcgt atgtacccag
     1261 aggacggagc cctcaaggga gagatcaagc aacgtctcaa gctcaaggac ggaggacact
     1321 acgacgccga ggtcaagacc acctacaagg ccaagaagcc agtccaactc ccaggtaagt
     1381 ttaaacatga ttttactaac taactaatct gatttaaatt ttcaggagcc tacaacgtca
     1441 acatcaagct cgacatcacc tcccacaacg aggactacac catcgtcgag caatacgagc
     1501 gtgccgaggg acgtcactcc accggaggaa tggacgagct ctacaaggga ttctccgaga
     1561 ccgtcgacct catgctcaac ctccaatcca acaaggaggg atccgtcgac ctcaagaacg
     1621 tctccgccgt cccaaaggag aagaccaccc tcaaggtaag tttaaacata tatatactaa
     1681 ctaaccctga ttatttaaat tttcaggacc catccaagcc accagccaag gcccaagtcg
     1741 tcggatggcc accagtccgt aactaccgta agaacatgat gacccaacaa aagacctcct
     1801 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     1861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gc
//
