LOCUS       sequence-390422-        5806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nucleolus-transport-enhanced Cre protein. Recombination-mediated 
            gene expression knockout..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5806)
  AUTHORS   Nava S, Palma W, Wan X, Oh JY, Gharib S, Wang H, Revanna JS, Tan M, 
            Zhang M, Liu J, Chen CH, Lee JS, Perry B, Sternberg PW
  TITLE     A cGAL-UAS bipartite expression toolkit for Caenorhabditis elegans 
            sensory neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Dec 19;120(51):e2221680120. doi: 
            10.1073/pnas.2221680120. Epub 2023 Dec 14.
  PUBMED    38096407
REFERENCE   2  (bases 1 to 5806)
  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."; date: "2023-12-19"; volume: "120(51):e2221680120. 
            doi"; pages: " 10.1073/pnas.2221680120. Epub 2023 Dec 14"
COMMENT     Sequence Label: pJR012
FEATURES             Location/Qualifiers
     source          1..5806
                     /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)"
     CDS             844..864
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             865..1893
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(2189..2217)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2965..2985
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        3551..3655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3656..3724,3725..4516)
                     /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(3874..3893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4687..5275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5176..5195
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5429..5446
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5563..5584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5599..5604,5605..5622,5623..5629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5637..5653
                     /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     5642..5664
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5661..5677
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5661..5677
                     /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 aaaaatgggc
      841 gcacccaaga agaagaggaa ggtgtccaat ttactgaccg tacaccaaaa tttgcctgca
      901 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
      961 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
     1021 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
     1081 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
     1141 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
     1201 tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
     1261 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     1321 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     1381 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     1441 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     1501 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     1561 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     1621 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     1681 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     1741 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     1801 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     1861 gcaatggtgc gcctgctgga agatggcgat tagggtaccg tagctgtctc atcctacttt
     1921 cacctagtta actgcttgtc ttaaaatcta tgcttctctt tagtatctaa aattttccta
     1981 gaagcttaca agtatataaa tggtctcttc tcaataaagg ttgtatattt attcatctta
     2041 ttgaatctgc catttcctcg tttttgcgag tttatatacc ttccaatttt ctttctattg
     2101 tattttcaac ttctaatttt aattcaggga aactgctgta ccggtagaaa aaatgctcga
     2161 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     2221 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     2281 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
     2341 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
     2401 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
     2461 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
     2521 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
     2581 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     2641 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     2701 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     2761 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     2821 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     2881 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     2941 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     3001 acatggcatg gatgaactat acaaatagga attcgctagc cggccataca agtaatccgg
     3061 atgatcgacg ccaacgtcgt tgaattttca aattttaaat actgaatatt tgtttttttt
     3121 cctattattt atttattctc tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat
     3181 ccaaatctaa acattttttt ttctctttcc gtctcccaat tcgtattccg ctcctctcat
     3241 ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg acgtgggggg
     3301 aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg acacacaata
     3361 tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat aaaaatatat
     3421 agaggaggaa ggaaaactag taaaaaataa gcaaagaaat taggcgaacg atgagaattg
     3481 tcctcgcttg ggcccaaaag gccttttttt tttggcgccg acgtcaggtg gcacttttcg
     3541 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3601 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3661 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3721 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3781 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3841 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3901 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3961 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4021 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4081 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4141 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4201 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4261 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4321 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4381 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4441 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4501 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4561 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4621 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4681 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4741 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4801 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4861 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4921 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4981 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5041 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5101 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5161 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5221 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5281 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5341 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5401 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     5461 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5521 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5581 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5641 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctgtaagt
     5701 ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt aaaaatggct
     5761 gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaat
//
