LOCUS       sequence-203285-        5848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dominant negative PKA cGAL effector for C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5848)
  AUTHORS   Wang H, Liu J, Yuet KP, Hill AJ, Sternberg PW
  TITLE     Split cGAL, an intersectional strategy using a split intein for 
            refined spatiotemporal transgene control in Caenorhabditiselegans.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Mar 26. pii: 1720063115. doi: 
            10.1073/pnas.1720063115.
  PUBMED    29581308
REFERENCE   2  (bases 1 to 5848)
  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: "Proc Natl 
            Acad Sci U S A. 2018 Mar 26. pii: 1720063115. doi: 
            10.1073/pnas.1720063115."
COMMENT     Sequence Label: pHW539 (15xUAS::kin-2a(G310D)::SL2::GFP::let-8...
FEATURES             Location/Qualifiers
     source          1..5848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..130
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    169..263
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    302..396
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     primer_bind     complement(2231..2259)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3007..3027
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        3593..3697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3698..3766,3767..4558)
                     /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     complement(3916..3935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4729..5317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5218..5237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5471..5488
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5605..5626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5641..5646,5647..5664,5665..5671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5679..5695
                     /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)."
     primer_bind     5684..5706
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5703..5719
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5703..5719
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcta gcccatgatt acgccaagct acgggcggag tactgtcctc cgagcggagt
       61 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
      121 gtcctccgag cggagactcc ctagcccatg attacgccaa gctacgggcg gagtactgtc
      181 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
      241 cgagcggagt actgtcctcc gagcggagac tccctagccc atgattacgc caagctacgg
      301 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg
      361 gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gactccctag gggcggccgc
      421 actgactggg ccggccaaaa atgcataagg ttttgctggc actgttcttt atctttctgg
      481 caccagcatc cgcactggca gtctccgaac cggcctgcag gtcgattttt tgcaaattac
      541 gagcgttgta gggggcggag cgataggtcc tataggtttt ggtatatcat cattcattca
      601 ttcattggta cattcattta cccaccttcc tctttctgag cttctctgga gttctgtgct
      661 tcctttttcc cttatcttta tactgtaatt tttaactttc aggcattgat tggcgcgcct
      721 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat
      781 aacatagaac attttcagga ggacccttgg ctagcgtcga cggtaccggt aaaaatgtcg
      841 ggtggaaacg aagaggatca gttggcacaa tgccaggcct atgtgcaacg acacaacatt
      901 cagcagcttg tcaaagaagc aattgttgtt ctgtgcatac acaaacccga caacccggtt
      961 ctcttcttga aggatcattt tgaaaaactc aacgaacaac gagcacagga aggaggaaat
     1021 ccggacgctg ctgatgatga cgacatcatt gtggagccac caaagagatc aggtggacgc
     1081 agaaccggaa tctctgccga accaatcaag gaggacgata ccgagtacaa gaaggttgtg
     1141 attccaaagg atgatgctac aagaaggtcg cttgagtcag ccatgaggaa gaacttgctc
     1201 tttgctcatt tggaggagga tgagcaaaaa accatgtacg atgccatgtt cccagttgag
     1261 aagtctgctg gagagaccat catcgaacag ggagaagaag gagataactt ctatgttatt
     1321 gataagggaa ccgttgatgt ctacgtcaat cacgagtacg ttctcaccat caacgagggt
     1381 ggatcatttg gagaacttgc tttgatctat ggaactccac gtgctgcaac cgtcattgcc
     1441 aagaccgacg tgaaattgtg ggccatcgac cgtctcacct accgtcgtat cttgatggga
     1501 tcagtaacaa agaagagaaa gatgtatgac gagttcctct ccaaggttca aatcttggcc
     1561 gatctcgacc agtgggagcg cgctaacgtg gctgatgctc ttgagagatg cgacttcgag
     1621 ccaggaactc atgtcgtcga gcaaggacaa cccggagacg aattcttcat cattctcgag
     1681 ggagaagcca atgtgttgca gaagagaagc gacgacgcac cattcgacgt tgtcggacat
     1741 cttggaatgt cggactattt cgacgaaatc gctcttcttc tcgaccgccc acgtgccgcc
     1801 accgttgttg ccaaaaccca tctcaagtgc atcaagctgg atcgtaaccg cttcgaacgt
     1861 gtcatgggac cagttcgtga gatcctgaag agagacgttt caaactacaa ctcatacgtc
     1921 aaactgatga cctaaggtac cgtagctgtc tcatcctact ttcacctagt taactgcttg
     1981 tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata
     2041 aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct
     2101 cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt
     2161 ttaattcagg gaaactgctg taccggtaga aaaaatgctc gagagtaaag gagaagaact
     2221 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     2281 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     2341 ttgcactact ggaaaactac ctgttccatg ggtaagttta aacatatata tactaactaa
     2401 ccctgattat ttaaattttc agccaacact tgtcactact ttctgttatg gtgttcaatg
     2461 cttctcgaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     2521 aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtaa
     2581 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcaggtg ctgaagtcaa
     2641 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
     2701 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
     2761 ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa
     2821 ctaatctgat ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt
     2881 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     2941 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     3001 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     3061 atacaaatag gaattcgcta gccggccata caagtaatcc ggatgatcga cgccaacgtc
     3121 gttgaatttt caaattttaa atactgaata tttgtttttt ttcctattat ttatttattc
     3181 tctttgtgtt ttttttcttg ctttctaaaa aattaattca atccaaatct aaacattttt
     3241 ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag
     3301 tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa
     3361 acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc
     3421 agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact
     3481 agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccaaa
     3541 aggccttttt tttttggcgc cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3601 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3661 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3721 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3781 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3841 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3901 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3961 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4021 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4081 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4141 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4201 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4261 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4321 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4381 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4441 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4501 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4561 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4621 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4681 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4741 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4801 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4861 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5101 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5341 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5401 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5461 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5521 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5581 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5641 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5701 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
     5761 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
     5821 ggatacgcta acaacttgga aatgaaat
//
