LOCUS       sequence-459058-        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For organ-specific expression of separated mCherry and UltraID in 
            the pharyngeal muscle of C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5723)
  AUTHORS   Liu J, Kulkarni A, Gao YQ, Urul DA, Hamelin R, Novotny BA, Long MJC,
            Aye Y
  TITLE     Organ-specific electrophile responsivity mapping in live C. elegans.
  JOURNAL   Cell. 2024 Dec 26;187(26):7450-7469.e29. doi: 
            10.1016/j.cell.2024.10.014. Epub 2024 Nov 5.
  PUBMED    39504959
REFERENCE   2  (bases 1 to 5723)
  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: 
            "10.1016/j.cell.2024.10.014"; journalName: "Cell"; date: 
            "2024-12-26- 26"; volume: "187"; issue: "26"; pages: "7450-7469.e29"
COMMENT     Sequence Label: pWormgate2-myo-2p-mCherry-P2A-flag-UltraID
FEATURES             Location/Qualifiers
     source          1..5723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1876..1932
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1939..1962
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3297..3319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3419..3438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3516..3534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3602..3706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3707..3775,3776..4567)
                     /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(3925..3944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4738..5326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5227..5246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5480..5497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5614..5635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5650..5655,5656..5673,5674..5680)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5688..5704
                     /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     5693..5715
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5712..5723,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5712..5723,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttcatttta tatctgagta gtatcctttg ctttaaatgt
       61 ccataaaact aattttataa tcaataaaac aacgtttgta aatcaactga gtttacaagt
      121 agagacattg agggatactt tcactatgct aaagtgataa tcgaccaaat aataacttca
      181 ctttggtatt tattcctgtc ttataaatgt tatgtatgaa ttaaattcat atgcatatgg
      241 ctcactctga caatttttaa taatcttcca gatcaatatt gactaccgat gcgggtggtc
      301 ttttgctttg aattctgctg aactttacac cccgaacagc aatgtgtgct tcagcctaaa
      361 aaaaagtaag tgtgttaatc agtgcccccg attcttcatt ttttgcccct ctctcccgtt
      421 tcgtcggcaa aagaagagaa aataaagata agtctcaaga taggttggta atcgctaaag
      481 tggttgtgtg gataagagta gcaaaatggc aggaagagca ctttgcgcgc acacactgta
      541 ctcattgttc tggataaaat tctctcgttg tttgccgtcg gatgtctgcc tctctgcatt
      601 gagccggctt cttcactatc tttagttaac ctaaaatgcc gtttcttttc tcgtatcccc
      661 actatcccgt tgaggttctc tgctctcttc gctccctacc gccagcgagc aactatccgt
      721 gggggcgcct tgctcggaag atggggggga agaaagaaga tttttgctat ttgcacttga
      781 gaaagagact tttcctgcgt cgatggttag agaacagtgt gcagacactt ttcagctacc
      841 tagaattaca attggatatc cccgcctccc aatccaccca cccagggaaa aagaagggct
      901 cgccgaaaat caaagttatc tccaggctcg cgcatcccac cgagcggttg acttctctcc
      961 accacttttc attttaaccc tcgatcgtca gacacagaaa tggctagcat ggtctcaaag
     1021 ggtgaagaag ataacatggc aattattaaa gagtttatgc gtttcaaggt gcatatggag
     1081 ggatctgtca atgggcatga gtttgaaatt gaaggtgaag gagaaggccg accatatgag
     1141 ggaacacaaa ccgcaaaact aaaggtaagt ttaaacatat atatactaac taaccctgat
     1201 tatttaaatt ttcaggtaac taaaggcgga ccattaccat tcgcctggga catcctctct
     1261 ccacagttca tgtatggaag taaagcttat gttaaacatc cggcagatat accagattat
     1321 ttgaaacttt cattcccgga gggttttaag tgggaacgcg taatgaattt tgaagacgga
     1381 ggagttgtta cagtgacgca agactcaagg taagtttaaa cagttcggta ctaactaacc
     1441 atacatattt aaattttcag cctccaagat ggagaattta tttataaagt caaacttcga
     1501 ggaacgaatt tcccctcgga tggacctgtt atgcagaaga agactatggg atgggaagct
     1561 tcaagtgaaa gaatgtaccc tgaagacggt gctcttaagg gagagattaa acaacgtctt
     1621 aaattgaaag atggaggaca ttacgatgct gaggtaagtt taaacatgat tttactaact
     1681 aactaatctg atttaaattt tcaggtgaag acaacttaca aagccaaaaa accagttcag
     1741 ctgccaggag cgtacaatgt taatattaaa ctggatatca cctcccacaa cgaggattac
     1801 actatcgttg agcaatatga aagagctgaa gggcggcact cgacaggtgg catggatgaa
     1861 ttgtataaga gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     1921 aaccctggac ctggcagcga ttataaagac gatgacgaca aggacttcaa gaacctgatc
     1981 tggctgaagg aggtggacag cacccaggag agactgaagg agtggaacgt gtcctacggc
     2041 accgccctgg tggccgacag acagaccaag ggcagaggcg gcccgggcag aaagtggctg
     2101 agccaggagg gcggcctgta cttcagcttc ctgctgaacc ccaaggagtt cgagaacctg
     2161 ctgcagctgc ccctggtgct gggcctgagc gtgagcgagg ccctggagga gatcaccgag
     2221 atccccttca gcctgaagtg gcccaacgac gtgtacttcc aggagaagaa ggtgagcggc
     2281 gtgctgtgcg agctgagcaa ggacaagctg atcgtgggca tcggcatcaa cgtgaaccag
     2341 agagagatcc ccgaggagat caaggacaga gccaccaccc tgtacgagat caccggcaag
     2401 gactgggaca gaaaggaggt gctgctgaag gtgctgaaga gaatcagcga gaacctgaag
     2461 aagttcaagg agaagtaaga gctccgcatc ggccgctgtc atcagatcgc catctcgcgc
     2521 ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct
     2581 gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt
     2641 agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc
     2701 gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa
     2761 aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga
     2821 aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga
     2881 ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt
     2941 gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt
     3001 ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt
     3061 tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt
     3121 gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct
     3181 atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat aactgtttct
     3241 actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3301 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3361 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     3421 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     3481 gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata
     3541 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3601 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3661 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3721 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3781 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3841 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3901 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3961 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4021 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4081 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4141 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4201 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4261 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4321 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4381 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4441 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4501 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4561 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4621 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4681 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4741 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4801 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4861 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4921 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4981 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5041 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5101 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5161 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5221 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5281 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5341 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5401 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5461 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     5521 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5581 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     5641 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     5701 acaatttcac acaggaaaca gct
//
