LOCUS       sequence-370755-        4006 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 4006)
  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 4006)
  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: pJJS010
FEATURES             Location/Qualifiers
     source          1..4006
                     /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(1821..1839)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1860..1876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1860..1876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1873..1895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1884..1900
                     /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(1908..1914,1915..1932,1933..1938))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1953..1974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2091..2108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2262..2850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3021..3812,3813..3881))
                     /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     3644..3663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3882..3986)
                     /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 tggtctccga gctcattaaa gaaaacatgc
      721 atatgaagct ctacatggag ggaaccgtca acaaccacca cttcaagtgc acctccgagg
      781 gagagggaaa gccatacgag ggaacccaaa ccatgcgtat caaggccgtc gagggaggac
      841 cactcccatt cgccttcgac atcctcgcca cctccttcat gtacggatcc aaggtaagtt
      901 taaacatata tatactaact aaccctgatt atttaaattt tcagaccttc atcaaccaca
      961 cccaaggaat cccagacttc ttcaagcaat ccttcccaga gggattcacc tgggagcgtg
     1021 tcaccaccta cgaggacgga ggagtcctca ccgccaccca agacacctcc ctccaagacg
     1081 gatgcctcat ctacaacgtc aaggtaagtt taaacagttc ggtactaact aaccatacat
     1141 atttaaattt tcagatccgt ggagtcaact tcccatccaa cggaccagtc atgcaaaaga
     1201 agaccctcgg atgggaggcc tccaccgaga ccctctaccc agccgacgga ggacttgagg
     1261 gacgtgccga catggccctc aagctcgtcg gaggaggaca cctcatctgc aacctcaagg
     1321 taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag accacctacc
     1381 gttccaagaa gccagccaag aacctcaaga tgccaggagt ctactacgtc gaccgtcgtc
     1441 ttgagcgtat caaggaggcc gacaaggaga cctacgtcga gcaacacgag gtcgccgtcg
     1501 cccgttactg cgacctccca tccaagctcg gacaccgtaa gggattctcc gagaccgtcg
     1561 acctcatgct caacctccaa tccaacaagg agggatccgt cgacctcaag aacgtctccg
     1621 ccgtcccaaa ggagaagacc accctcaagg taagtttaaa catatatata ctaactaacc
     1681 ctgattattt aaattttcag gacccatcca agccaccagc caaggcccaa gtcgtcggat
     1741 ggccaccagt ccgtaactac cgtaagaaca tgatgaccca acaaaagacc tcctccgcgg
     1801 tggagctcca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
     1861 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1921 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1981 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2041 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2101 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2161 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2701 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2881 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3061 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3121 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3181 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3241 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3301 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3361 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3421 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3481 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3541 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3601 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3661 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3721 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3781 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3841 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3901 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3961 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgc
//
