LOCUS       sequence-97553-e        6805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zf1::yfp fusion with floxed unc-119(+) within an intron of yfp. For 
            creating zf1::yfp knock-ins using unc-119 as a marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6805)
  AUTHORS   Armenti ST, Lohmer LL, Sherwood DR, Nance J
  TITLE     Repurposing an endogenous degradation system for rapid and targeted 
            depletion of C. elegans proteins.
  JOURNAL   Development. 2014 Dec;141(23):4640-7. doi: 10.1242/dev.115048. Epub 
            2014 Nov 5.
  PUBMED    25377555
REFERENCE   2  (bases 1 to 6805)
  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: 
            "10.1242/dev.115048"; journalName: "Development"; date: "2014-12"; 
            volume: "141"; issue: "23"; pages: "4640-7"
COMMENT     Sequence Label: pJN601
FEATURES             Location/Qualifiers
     source          1..6805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(154..182)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    522..555
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2525..2536
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    2685..2718
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3127..3147
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4121..4143)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4243..4262
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4340..4358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4426..4530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4531..4599,4600..5391)
                     /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(4749..4768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5562..6150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6051..6070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6304..6321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6438..6459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6474..6479,6480..6497,6498..6504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6512..6528
                     /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     6517..6539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6536..6552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6536..6552
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acagaataca aaacgcgact ttgtgatgcg ttccgccgtg aaggatactg cccgtacaac
       61 gacaattgca catatgctca cggacaagat gagctgagag ttccgagggt accggtagaa
      121 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
      181 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
      241 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
      301 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
      361 actttcggtt atggtcttca atgcttcgcc agatacccag atcatatgaa acagcatgac
      421 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
      481 gacgggaact acaagacacg taagtttaaa cagttcggta cataacttcg tataatgtat
      541 gctatacgaa gttatgccta gttctagaca ttctctaatg aaaaaatctt tcagttgaaa
      601 ttgaaaatga gttaaagttg gagtttttat tgaaaacaga tttccgtgtg attagtgttt
      661 ttagcgagtg tgacaggaca gcgaaaaaat atagaaacaa ggggggaact gaaaagctta
      721 ggaatgcatt gaacatgaga aggggaaggg gaaggaacaa actagacagg aattattgga
      781 atttaatcac atttggagtt ttttttctat tcgacagaat aattatccag aacatttttg
      841 tattaaatat ttatgcatca tatgagtagt cggctttgtt gtgcatgacg agtttgttat
      901 cgacgaaata gaagctgtca gaacgagtct cgtttggatt gttgatcatg tcgtccactg
      961 aaaaagagat tagtcttttg aattgtactt tttagaataa tgactcactg agttgttgag
     1021 agagttgagg gaactcatag atatgttcac agttgtttcg tgaattcgga atacagaatc
     1081 cgaattcaaa gtcaaaacac ttcagaaggc gatctttgaa gaagtgacgt tcgatcattc
     1141 ggaaatgatg gattgggatg tctcctactt tgaattccac agttgctccg accgttttga
     1201 gtttcaaaaa gtttggagca aatctataac gcacgtatct tgccgactcc tgtggcgact
     1261 catcattctc ctgatcgttc tccggtttgg cgatctcgaa aagcacttgc tcagtgtcca
     1321 gatcacggat ttggaacttg gtgaactcga tgttatagat gttcgcagat ggggagcata
     1381 agaatcctaa atttatgttt taaactgaaa tccaaaggga gcaagatacc ttgagtgatt
     1441 cccggaagtg ctaaaacgtc gttcggagtg atttgagctt tcttcgcaag ctccgattcc
     1501 gttgtgattc cttgttcggt gcttggtggt ggccgtggca tctggaaata tggaaaagtt
     1561 caacaaaaag aaaagagaaa agaatgaaat cggatatcaa gagttagttg agcggtttct
     1621 ctagttttct gagtctcacc tgcgacggga aggtcgccga gccgggtgga atcgatcgtt
     1681 gttgctcggc tttcatatcg gtttggttgg aagcggctga aaacggaaag aagtggaaga
     1741 aggaaaagag tgtggtgtga caggaaaatg gtaattagag ggtgccaaat aaccagctat
     1801 attttgtttt tttttgaaaa catttttaaa aagaaaaata cgataatgat atcagatgga
     1861 tttccggaaa actggtatga aaaatttcaa cctttttgag tacatgtaat caaaatacac
     1921 tttgtaaatt atcattttta ttgaaactcc accatttttc tatttataac gctaataatt
     1981 tgaaaaagaa acctattgcg aaccgcgggg tgaatcccaa aaacgaatgc gttttggtgg
     2041 agtgattgat tcgaatcgaa gaagaaaaag aagaagacgt ggaatagaga gctcactctt
     2101 aaccgagcag cacacaccga cagaaaaaaa aatgaaatga atgagggtct tcttcttctt
     2161 cttcttcgaa tgattgacag aaatgggaaa aagaggaaga ttgagaaggg aaaaaggaag
     2221 gagaaaagaa gcagaagaag acgtcagaga ggagaggaac gagcggaaaa gcagcgggcg
     2281 caagtcatag aagtagcaga gctggggaga agaagacact atccaagaaa ggaatgacga
     2341 gagagtatgc aaaggggtat agggtgcaga cagaatagga acagaataac agatgatgag
     2401 ccaagaagag ttgaaaaggg cgatgaattt gtcatgtaac ttaatttggg tcaatttgag
     2461 catgatgaat tgaaatcatc ccttgttggg agttaataac cggtttgtta tcagaaaccc
     2521 tgtaatagaa gggcgcccta actttgagcc aattcatccc ggtttctgtc aaatatatca
     2581 aaaagtggtc aactgacaaa ttgtttttga tattataata aacattttat ccgttaacaa
     2641 ttttcgaata ctttttacaa ggacttggat aaattggctc aaagataact tcgtataatg
     2701 tatgctatac gaagttatta actaaccata catatttaaa ttttcaggtg ctgaagtcaa
     2761 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga
     2821 tggaaacatt cttggacaca aattggaata caactataac tcacacaatg tatacatcat
     2881 ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa
     2941 ctaatctgat ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt
     3001 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     3061 agacaaccat tacctgtcct accaatctgc cctttcgaaa gatcccaacg aaaagagaga
     3121 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     3181 atacaaatag cattcgtaga attccaactg agcgccggtc gctaccatta ccaacttgtc
     3241 tggtgtcaaa aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa
     3301 ctaacgagta atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca
     3361 attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt
     3421 attatcaaaa aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta
     3481 acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat
     3541 tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta
     3601 cacttcttat gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg
     3661 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     3721 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     3781 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     3841 ggggtttccc ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac
     3901 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     3961 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     4021 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     4081 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4141 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4201 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4261 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4321 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4381 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4501 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4561 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4621 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4681 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4741 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4801 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4861 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4921 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4981 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5041 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5101 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5161 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5221 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5281 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5341 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5401 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5701 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5941 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6241 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6301 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6361 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6421 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6481 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6541 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     6601 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     6661 ctaacaactt ggaaatgaaa taagcttgca tgcctgcagg tcgactctag aggatccccg
     6721 ggattggcca aaggacccaa aggtatgttt cgaatgatac taacataaca tagaacattt
     6781 tcaggaggac ccttggaggg taccg
//
