LOCUS       sequence-141613-        5580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3XFLAG::GR LBD::eGFP from the hsp16.48 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  AUTHORS   Monsalve GC, Yamamoto KR, Ward JD
  TITLE     A New Tool for Inducible Gene Expression in Caenorhabditis elegans.
  JOURNAL   Genetics. 2019 Feb;211(2):419-430. doi: 10.1534/genetics.118.301705.
            Epub 2018 Nov 30.
  PUBMED    30504365
REFERENCE   2  (bases 1 to 5580)
  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"; doi: 
            "10.1534/genetics.118.301705"; journalName: "Genetics"; date: 
            "2019-02"; volume: "211"; issue: "2"; pages: "419-430"
COMMENT     Sequence Label: pJW1241
FEATURES             Location/Qualifiers
     source          1..5580
                     /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        730..1034
                     /label=hsp16-48 promoter
                     /note="promoter for the C. elegans heat shock protein 16-48
                     gene (Russnak and Candido, 1985)"
     CDS             1048..1113
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1120..1140
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2005..2307,2361..2435,2487..2606,2658..2873)
                     /codon_start=1
                     /product="monomeric enhanced GFP, optimized with synthetic 
                     introns for expression in C. elegans"
                     /label=mEGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITHGMDELYK"
     CDS             2880..2954
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="MSRRRKANPTKLSENAKKLAKEVEN"
     terminator      2964..3287
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     promoter        complement(3395..3413)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3434..3450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3434..3450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3447..3469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3458..3474
                     /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(3482..3488,3489..3506,3507..3512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3527..3548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3665..3682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3916..3935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4595..5386,5387..5455))
                     /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     5218..5237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5456..5560)
                     /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 tgggtaccgc
      661 tggacggaaa tagtggtaaa gtgacatgat tgtagtttga agatttcaca attagagtga
      721 atgttgtttg gttcggtttt gtcactgtat ttatactcat ttccaccttt ttctagaaca
      781 ttcgagctgc ttcttacaaa aggaggcgac tcacattcag aacattgaga aatagtgtgc
      841 gactgaagaa acccagatac tttttcaatc tgcgtctctt tgcacctatg gggtgtattt
      901 tgaaatgaat gcatctagga ccttctagaa cattctaaac ggctgcagga tacgggtata
      961 taagccaatc gtgttcagag gaaaccaata cactttgttc aagtgcttac tgttcattct
     1021 ctaaacttca agaactagaa aaaaatggac tacaaggacc acgacggaga ctacaaggac
     1081 cacgacatcg actacaagga cgacgacgac aagatggccc caaagaagaa gcgtaaggtc
     1141 ggaatccacg gagtcccagc cgccattcag caggccacta caggagtctc acaagaaacc
     1201 tctgaaaatc ctggtaacaa aacaatagtt cctgcaacgt taccacaact cacccctacc
     1261 ctggtgtcac tgttggaggt tattgaacct gaagtgttat atgcaggata tgatagctct
     1321 gttccagact caacttggag gatcatgact acgctcaaca tgttaggagg gcggcaagtg
     1381 attgcagcag tgaaatgggc aaaggcaata ccaggtttca ggaacttaca cctggatgac
     1441 caaatgaccc tactgcagta ctcctggatg tttcttatgg catttgctct ggggtggaga
     1501 tcatatagac aatcaagtgc aaacctgctg tgttttgctc ctgatctgat tattaatgag
     1561 cagagaatga ctctaccctg catgtacgac caatgtaaac acatgctgta tgtttcctct
     1621 gagttacaca ggcttcaggt atcttatgaa gagtatctct gtatgaaaac cttactgctt
     1681 ctctcttcag ttcctaagga cggtctgaag agccaagagc tatttgatga aattagaatg
     1741 acctacatca aagagctagg aaaagccatt gtcaagaggg aaggaaactc cagccagaac
     1801 tggcagcggt tttatcaact gacaaaactc ttggattcta tgcatgaagt ggttgaaaat
     1861 ctccttaact attgcttcca aacatttttg gataagacca tgagtattga attccccgag
     1921 atgttagctg aaatcatcac caatcagata ccaaaatatt caaatggaaa tatcaaaaaa
     1981 cttctgtttc atcaaaagct gcagatgtcc aagggagagg agctcttcac cggagtcgtc
     2041 ccaatcctcg tcgagctcga cggagacgtc aacggacaca agttctccgt ctcaggagag
     2101 ggagagggag acgccaccta cggaaagctc accctcaagt tcatctgcac caccggaaag
     2161 ctcccagtcc catggccaac cctcgtcacc accttcactt acggagtcca atgcttctcc
     2221 cgttacccag accacatgaa gcgtcacgac ttcttcaagt ccgccatgcc agagggatac
     2281 gtccaagagc gtaccatctt cttcaaggta agtttaaaca ttaattaata ctaactaacc
     2341 ctgattattt aaattttcag gacgacggaa actacaagac ccgtgccgag gtcaagttcg
     2401 agggagacac cctcgtcaac cgtatcgagc tcaaggtaag tttaaacagt tcggtactaa
     2461 ctaaccatac atatttaaat tttcagggaa tcgacttcaa ggaggacgga aacatcctcg
     2521 gacacaagct cgaatacaac tacaactccc acaacgtcta catcatggcc gacaagcaaa
     2581 agaacggaat caaggtcaac ttcaaggtaa gtttaaacat gattttacta actaactaat
     2641 ctgatttaaa ttttcagatc cgtcacaaca tcgaggacgg atctgtccaa ctcgccgacc
     2701 actaccaaca aaacacccca atcggagacg gaccagtcct cctcccagac aaccactacc
     2761 tctccaccca atccgccctc tccaaggacc caaacgagaa gcgtgaccac atggtcctca
     2821 aggagttcgt caccgctgcc ggaatcaccc acggaatgga cgagctctac aagctgcaga
     2881 tgtcccgtcg tcgtaaggcc aacccaacca agctctccga gaacgccaag aagctcgcca
     2941 aggaggtcga gaactaggaa ttcatgcaag atcctttcaa gcattccctt cttctctatc
     3001 actcttcttt ctttttgtca aaaaattctc tcgctaattt atttgctttt ttaatgttat
     3061 tattttatga ctttttatag tcactgaaaa gtttgcatct gagtgaagtg aatgctatca
     3121 aaatgtgatt ctgtctgatg tactttcaca atctctcttc aattccattt tgaagtgctt
     3181 taaacccgaa aggttgagaa aaatgcgagc gctcaaatat ttgtattgtg ttcgttgagt
     3241 gacccaacaa aaagaggaaa ctttattgtg ccgccaagaa aaaagtctca ttgcctaaga
     3301 gaaacatatg tggttggaaa tcttccagat cgtggatcac tagtatgtgg ttggaaatct
     3361 tccagatcgt ggatcagagc tccagctttt gttcccttta gtgagggtta atttcgagct
     3421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
//
