LOCUS       sequence-458991-        6419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For T-REX experiments of asp-2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  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 6419)
  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: pCS2+8-asp-2-flag-tev-halo-his
FEATURES             Location/Qualifiers
     source          1..6419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1429..1452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1459..1479
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1486..2376
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2386..2403
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2471..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2495..2514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2549..2568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2718..2738)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2718..2736)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2757..2773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2757..2773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2770..2792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2781..2797
                     /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)."
     promoter        complement(join(2805..2811,2812..2829,2830..2835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2850..2871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2988..3005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3159..3747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3239..3258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3918..4709,4710..4778))
                     /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     4541..4560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4779..4883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4909..5364)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5046..5067)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5258..5277
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5435..6419
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccactagt ggcgcgccat taattaaagg ccggccagcg
      121 atcgccggac ccaccatgca gaaccttttt ttgttgcttg cccttgtggc agcagctctc
      181 gctgcagttg tgcaagttcc actcgtgaaa attgagccat acagaaaccg attgatccgt
      241 gaaggaagat gggttcagta cagaaaggat cgtgagattc gtcgtttcat gatgaacaag
      301 caatcaaatg atatgtctgt tggacaatac gtcaacgatt acgaagatga agcatacgtc
      361 ggaaacatca ccattggaac tccacaacaa caattcaagg ttattcttga tactggatca
      421 tccaatcttt ggatcccaga catcacatgt ggaactaagc cagaaaattg ctcaaccgtt
      481 ccagcttgcc gtggaattat ttgccaattc gaatgcgacg accaagcttg ctgtggagct
      541 ggagcgaact acaccgattc ttgcttctac caaaacaagt ttgacgcttc caaatcttcc
      601 tcatacaaga ccaatggaag aaacttcatt attgagtatg gaacaggatc cgccagagga
      661 ttccttggac aggacactat caccttcgga ggaattggag aaccacaact cgctgttcca
      721 aacactgttt tcggacaagc cacctcactt gctgccttct tcgagggaca acctcttgat
      781 ggaatccttg gacttgcgtt caaatcgatt gctgttgatc aaatcactcc accattcatc
      841 aacgccatca accaaaacct tgttgatctt ccacttttca ccgtcttcct tgagcatgaa
      901 ggagatcaaa acggagttca aggaggagtc tacacttatg gaggaattga tacaaagaac
      961 tgtggtccag ttatcgctta tcaaccattg tcctctgcca cttattacca attcaagatg
     1021 agtgctattg gatctggaag ctatcactct agcaagggat ggcaagtcat ctccgacacc
     1081 ggaacttctc tcatcggagg accaaaggcc tacgtttctg ctattgctga cgctgttgga
     1141 gccacatgga gagatgatta cggtgtatac atccttccat gctctgcaaa gatcaacact
     1201 cttgatatca ccattggaaa ccaagtatac aatattgact catccaacac cattattcca
     1261 ctcggagatg gaagcaacaa ttgcatctac gccatcttcc cattcagcag tggaggtttt
     1321 ggaccatcat ggattcttgg agatccattc attcgtcaat attgtaacat ctatgatgtt
     1381 ggaactcaac gtgttggatt tgctaagtcc ctccaatcga aatcagggga ttataaagac
     1441 gatgacgaca agtcagggga aaacttgtat ttccagggct cagggatggc agaaatcggt
     1501 actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc
     1561 gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc gacctcctcc
     1621 tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat tgctccagac
     1681 ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga cgaccacgtc
     1741 cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct ggtcattcac
     1801 gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg cgtcaaaggt
     1861 attgcattta tggagttcat ccgccctatc ccgacctggg acgaatggcc agaatttgcc
     1921 cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat catcgatcag
     1981 aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag
     2041 atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact gtggcgcttc
     2101 ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac
     2161 atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac cccaggcgtt
     2221 ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg
     2281 gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat cggcagcgag
     2341 atcgcgcgct ggctgtcgac gctcgagatt tccggcggca gcagccatca tcatcatcat
     2401 catgggtcag ggtaataatc gaattcaagg cctctcgagc ctctagaact atagtgagtc
     2461 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2521 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2581 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2641 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg
     2701 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     2761 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg
     2821 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2881 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2941 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3001 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3061 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4861 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     4921 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4981 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5041 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5101 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5161 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5221 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5281 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5341 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     5401 ggcgataggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
     5461 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     5521 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     5581 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
     5641 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     5701 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     5761 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     5821 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     5881 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     5941 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     6001 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     6061 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     6121 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     6181 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
     6241 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     6301 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     6361 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
