LOCUS       sequence-459003-        6826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For organ-specific expression of separated mCherry and UltraID in 
            the intestine of C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6826)
  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 6826)
  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-ges-1p-mCherry-P2A-flag-UltraID
FEATURES             Location/Qualifiers
     source          1..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2979..3035
                     /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             3042..3065
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4400..4422)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4522..4541
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4619..4637)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4705..4809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4810..4878,4879..5670)
                     /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(5028..5047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5841..6429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6330..6349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6583..6600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6717..6738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6753..6758,6759..6776,6777..6783)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6791..6807
                     /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     6796..6818
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6815..6826,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6815..6826,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgtgctta aagagaagac tgttagccac tcagccactt
       61 cagccatatc gaaacagtca tattggctct ctttcagatc acttaaaact ccgaactatg
      121 atgacgaaaa aatgttttct tgaattttca ttttattgtg aaattgaaaa ctctgcatag
      181 cccgcagtca cttgaaagca catttaagat ggaatcaaat ggtaagaaag ctcgtgtaat
      241 caggagattt tatccggggt atgccgagga atatatggtg cactcgcatt agctggcatt
      301 ttttgtaatt atcggatttc tacctttttc ttaatctagt ttgttatcat tgtccatcat
      361 ttacaaccaa aaatttgcgt catttacaac caaaaattga tctgacaatt atcataattc
      421 ggtttttatt ttcagaagaa acgctccgat gtgatcctat agtgaatata cttttccatc
      481 ctctctgaaa taaacaatta tgaagaatgc atcaatttgc attcgaaata actgccgtat
      541 tttgtctatt ggtatggctg cagtaccatt tttcttcgcg tctttgcgca gccttcgggc
      601 gctaccaata aggctaagcc ggggtaagtt atttcagaat tctaatcaac taatctaagt
      661 ctcatacatc attgtcaagt gacgcatttt tttaatttct atgttaactc gactttaata
      721 ttcttttctg gtttcaccag aaaaatgctg gactgaaaaa ttgaatttcc cgcgatttgg
      781 gaaaaaccgc agtttaagct tttggcatga atacagtgaa cacaggaaca aaaaattgac
      841 ttgttatttt gagtttatat taaatgttaa ttttgcatct catattttca caaaacttgt
      901 ttgcttgaaa aatgtgcaaa taacaagtga acatacatca aggtttatgc agatcagcac
      961 tgatgcaact gatagcaaaa ctgataaggg tcaaaatttc agtggccagc acaaacacat
     1021 acacattcgt agttcgtttt gtgtttcgtg tatttagcat ttatttttca attccggcaa
     1081 tttgccgatt tgccggaaat tttgattttt ggcaaattgc cagtttgccg tttgccgaac
     1141 atcaatttgc cgaaattttt tagagggatc tttataagac ggaaacactt aaaattgtgc
     1201 ttttttggaa tttttttcaa tgtttcaata gaattagctt acatttaaaa ttagatgtag
     1261 gaacgtttat aggacgctta caattttgcc gattaaaatc gaagttctga acttttcaaa
     1321 aaatgtgcaa aatttttttt gacaaatttg acaaatcggc cggtttgtcg attgtccgga
     1381 atttttcagt ttcggcgatt taccggtttg ccggaaaaca tcgtttgcca cccacacaca
     1441 cataaagaga cgaaacaaca caccatcttc aaaaaatttg tgaatcttca tttccacgtg
     1501 gtttgcaggg tttcccatcc accccactct atctctccaa gttgaatttt ggcaaaattg
     1561 accacacacc gatcaaaaac tattgtgtgt gtgtgtattt gtgtcttcaa taattcctgt
     1621 ctgtgtctcg cactatctat tgactacaag aacgtgatcg cagctctacg aaacctttac
     1681 tttattttag aaatattttt tcatgctatt agcaaaaaaa aagttttttt ttgtctaagt
     1741 acacttaaaa ttttccaaaa tttttaagat ttttcaaaat gtgcgatcaa agatctgaca
     1801 aaatgccaaa attttgaaga ggaaatttag tcggcttcca aaattataag tggcagaaat
     1861 tgagcaattg tcactttttg acagtaaata gaaaattttt tgaaaaattt tgaaacgctt
     1921 tattttgtta cttggtcatt ttggcaccat aggagtagtt tttaacattt tccccactgg
     1981 cgctacttca ttttaaactt ttccaaagat tttaattttt aattttaaaa aatttcatat
     2041 aattcgtctg cgtcttacag tttcaggctt caaaaatcta ttacatatct tatctttgaa
     2101 ttcaggctag catggtctca aagggtgaag aagataacat ggcaattatt aaagagttta
     2161 tgcgtttcaa ggtgcatatg gagggatctg tcaatgggca tgagtttgaa attgaaggtg
     2221 aaggagaagg ccgaccatat gagggaacac aaaccgcaaa actaaaggta agtttaaaca
     2281 tatatatact aactaaccct gattatttaa attttcaggt aactaaaggc ggaccattac
     2341 cattcgcctg ggacatcctc tctccacagt tcatgtatgg aagtaaagct tatgttaaac
     2401 atccggcaga tataccagat tatttgaaac tttcattccc ggagggtttt aagtgggaac
     2461 gcgtaatgaa ttttgaagac ggaggagttg ttacagtgac gcaagactca aggtaagttt
     2521 aaacagttcg gtactaacta accatacata tttaaatttt cagcctccaa gatggagaat
     2581 ttatttataa agtcaaactt cgaggaacga atttcccctc ggatggacct gttatgcaga
     2641 agaagactat gggatgggaa gcttcaagtg aaagaatgta ccctgaagac ggtgctctta
     2701 agggagagat taaacaacgt cttaaattga aagatggagg acattacgat gctgaggtaa
     2761 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggtg aagacaactt
     2821 acaaagccaa aaaaccagtt cagctgccag gagcgtacaa tgttaatatt aaactggata
     2881 tcacctccca caacgaggat tacactatcg ttgagcaata tgaaagagct gaagggcggc
     2941 actcgacagg tggcatggat gaattgtata agagcggagc tactaacttc agcctgctga
     3001 agcaggctgg agacgtggag gagaaccctg gacctggcag cgattataaa gacgatgacg
     3061 acaaggactt caagaacctg atctggctga aggaggtgga cagcacccag gagagactga
     3121 aggagtggaa cgtgtcctac ggcaccgccc tggtggccga cagacagacc aagggcagag
     3181 gcggcccggg cagaaagtgg ctgagccagg agggcggcct gtacttcagc ttcctgctga
     3241 accccaagga gttcgagaac ctgctgcagc tgcccctggt gctgggcctg agcgtgagcg
     3301 aggccctgga ggagatcacc gagatcccct tcagcctgaa gtggcccaac gacgtgtact
     3361 tccaggagaa gaaggtgagc ggcgtgctgt gcgagctgag caaggacaag ctgatcgtgg
     3421 gcatcggcat caacgtgaac cagagagaga tccccgagga gatcaaggac agagccacca
     3481 ccctgtacga gatcaccggc aaggactggg acagaaagga ggtgctgctg aaggtgctga
     3541 agagaatcag cgagaacctg aagaagttca aggagaagta agagctccgc atcggccgct
     3601 gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac
     3661 atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac
     3721 ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt
     3781 agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc
     3841 cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt
     3901 ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc
     3961 atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca
     4021 tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca
     4081 agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt
     4141 gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg
     4201 caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat
     4261 aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca
     4321 atataccaaa cataactgtt tctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     4381 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4441 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4501 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4561 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     4621 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4801 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4861 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4921 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4981 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5041 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5101 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5161 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5221 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5281 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5341 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5401 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5461 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5521 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5581 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5641 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5701 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5761 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6601 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6661 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6721 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6781 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagct
//
