LOCUS       sequence-459536-        6026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For organ-specific expression of GFP-Halo in the pharyngeal muscle 
            of C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  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 6026)
  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-gfp-halo
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1039..1067)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1815..1835
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1888..2778
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(3600..3622)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3722..3741
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3819..3837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3905..4009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4010..4078,4079..4870)
                     /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(4228..4247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5041..5629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5530..5549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5783..5800
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5917..5938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5953..5958,5959..5976,5977..5983)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5991..6007
                     /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     5996..6018
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6015..6026,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6015..6026,1..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 gagtaaagga
     1021 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     1081 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     1141 aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa catatatata
     1201 ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt ctgttatggt
     1261 gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt caagagtgcc
     1321 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1381 acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt ttcaggtgct
     1441 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     1501 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     1561 tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa catgatttta
     1621 ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa cattgaagat
     1681 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     1741 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     1801 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     1861 gatgaactat acaaatcagg gtcaggcatg gcagaaatcg gtactggctt tccattcgac
     1921 ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat
     1981 ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc
     2041 atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa
     2101 tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc
     2161 atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg
     2221 ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc
     2281 atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc
     2341 ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt
     2401 acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag
     2461 ccgttcctga accctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc
     2521 gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag
     2581 tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa
     2641 gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg
     2701 aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg
     2761 acgctcgaga tttccggcta agagctccgc atcggccgct gtcatcagat cgccatctcg
     2821 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     2881 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     2941 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     3001 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     3061 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     3121 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     3181 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     3241 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     3301 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     3361 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     3421 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     3481 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     3541 tctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3601 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3661 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     3721 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     3781 tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt
     3841 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5701 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5761 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5821 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5881 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5941 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6001 ataacaattt cacacaggaa acagct
//
