LOCUS       sequence-459537-        7547 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For organ-specific expression of GFP-Halo in the body-wall muscle 
            and neighboring vulval muscle cells of C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  TITLE     Organ-specific electrophile responsivity mapping in live C. elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7547)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pWormgate2-myo-3p-gfp-halo
FEATURES             Location/Qualifiers
     source          1..7547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2560..2588)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3336..3356
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3409..4299
                     /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(5121..5143)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5243..5262
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5340..5358)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5426..5530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5531..5599,5600..6391)
                     /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(5749..5768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6562..7150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7051..7070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7304..7321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7438..7459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7474..7479,7480..7497,7498..7504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7512..7528
                     /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     7517..7539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7536..7547,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7536..7547,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttagtgatt atagtctctg ttttcgttaa ttttgaattt
       61 tgcttgataa ggctgcaaca aagatcaggt tgacatatat tttcagtaat ttattttaac
      121 ctgtactcta tcactgccgg ctataataag ttcttgaata aaataatttt cccgacaaaa
      181 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      241 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      301 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      361 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      421 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      481 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      541 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      601 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      661 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      721 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      781 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      841 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      901 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      961 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
     1021 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
     1081 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1141 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1201 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1261 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1321 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1381 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1441 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1501 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1561 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1621 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1681 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1741 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1801 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1861 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1921 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1981 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     2041 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     2101 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2161 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2221 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2281 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2341 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2401 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2461 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2521 gaagctagca tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     2581 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     2641 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     2701 gtaagtttaa acatatatat actaactaac cctgattatt taaattttca gccaacactt
     2761 gtcactactt tctgttatgg tgttcaatgc ttctcgagat acccagatca tatgaaacgg
     2821 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
     2881 aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa ctaaccatac
     2941 atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     3001 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     3061 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     3121 gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca gaacttcaaa
     3181 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     3241 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     3301 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     3361 gctgggatta cacatggcat ggatgaacta tacaaatcag ggtcaggcat ggcagaaatc
     3421 ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac
     3481 gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc
     3541 tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca
     3601 gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac
     3661 gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt
     3721 cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa
     3781 ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt
     3841 gcccgcgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat
     3901 cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc
     3961 gagatggacc attaccgcga gccgttcctg aaccctgttg accgcgagcc actgtggcgc
     4021 ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa
     4081 tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc
     4141 gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct
     4201 gtggacatcg gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc
     4261 gagatcgcgc gctggctgtc gacgctcgag atttccggct aagagctccg catcggccgc
     4321 tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa
     4381 catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa
     4441 cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg
     4501 tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc
     4561 ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt
     4621 tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat
     4681 catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc
     4741 atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc
     4801 aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt
     4861 tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     4921 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     4981 taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc
     5041 aatataccaa acataactgt ttctactagt cggccgtacg ggccctttcg tctcgcgcgt
     5101 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5161 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5221 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     5281 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     5341 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5521 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5581 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5641 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5701 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5761 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5821 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5881 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5941 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6001 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6061 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6121 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6181 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6241 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6301 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6361 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6421 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6481 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6541 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6601 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6661 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7021 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7081 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7141 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7201 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7261 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7321 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7381 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7441 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7501 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
