LOCUS       sequence-458989-        6743 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For T-REX experiments of aex-5..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  TITLE     Organ-specific electrophile responsivity mapping in live C. elegans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6743)
  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            "Unpublished"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: pCS2+8-aex-5-flag-tev-halo-his
FEATURES             Location/Qualifiers
     source          1..6743
                     /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             1753..1776
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1783..1803
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1810..2700
                     /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             2710..2727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2795..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2819..2838)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2873..2892
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3042..3062)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3042..3060)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3081..3097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3081..3097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3094..3116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3105..3121
                     /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(3129..3135,3136..3153,3154..3159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3174..3195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3312..3329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3483..4071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3563..3582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4242..5033,5034..5102))
                     /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     4865..4884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5103..5207)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5233..5688)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5370..5391)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5582..5601
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5759..6743
                     /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 ccaccatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt
      181 gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac
      241 gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaagct tcaatcatac
      301 accgatgtgt acgttggtcg ccgacttcgt cgtcgaaagc gtggaattga agatgagatt
      361 gtggcggaga tgatgctaag tcaacaagtg caatttattg aaaaacttca aggatttaga
      421 agatacaaaa gagcacccat gacaaacaaa ggatatcctg ttcatgtgtg gaacctaaca
      481 ccttcacttt acatccggga agcttgggaa gatgggttca acgcaagccg ggttactgta
      541 gcagtggtgg atgatggtgt tgatatcaaa cacgttgatt tgaagagtgc attctctccg
      601 cgcgtttcct ttgattttgt gcgattcggc gatctgccaa caccgaaaaa ttctaaagag
      661 tttgagcatg gaacccaatg tgctggattg gtagcaatgg aaggacaaca atgcggcctt
      721 ggtgtaggtc atggtgcaac actgggtgct ataaagcttt tgggtcagga cttcttgaac
      781 gacgcgttgg aaggagatgc cttggcattt cagaaagact tgattgacat ctactcagtc
      841 agctggggac ccaaagacga tggtaaaagt gcggagaagc cggcaaagtt cacagaagaa
      901 gctattaaga atggagcatt gcacggaaga aatggaaaag gaaacatatt tgtgtgggcg
      961 agtggtaatg gtggagtcaa tggggataat tgtgcatatg atgggtatgt tagcaatgag
     1021 tatacgctga gctttggagt cattgatgcg agtggagcac ctgctgcata cggagaagga
     1081 tgttcgtcgg ttcttgcagc agtttctggt ggggatgcaa tgattcaaac cactggccta
     1141 gaatccacgt gttcctcaat atccggatcg tcggcttctg cggcaattgc atcaggaatc
     1201 atttcattgg tattggatgc taatcccaca ctatcccaac gtgacattca acacctcatt
     1261 gctcgcacct ccaatgcaag tgcaatccgt gacgtggaat tgtatgaaaa ttctgctggc
     1321 ctaaatttcc acccaaaagt tggattcggt ctactgaatg cacagaaact cgtggtaatg
     1381 gctgccacgt gggagaatgt ggcacctcaa gtgacttgcg agaaaatgaa tctagcgaat
     1441 ggaataatcg ataattcgga ttgtgacgtc acaaaagtag aacgtgtaat tgtctccgga
     1501 tctatcatcc atccacatcg aggacaagtt caaatcagat tggaatcgcc acggggcaca
     1561 atctccgaac tgcttccact tcgtcccaaa gacacatcgc gagatcttct cgactggaac
     1621 tttgtatccg tcaacttttt tggagagaat tcacgtggta tctggaagtt gcatgtgaca
     1681 agtgaagagg atgacgtgga tttcagagtt gagatgaaaa tgttcaaagt ggtgggaaca
     1741 atgtcatcag gggattataa agacgatgac gacaagtcag gggaaaactt gtatttccag
     1801 ggctcaggga tggcagaaat cggtactggc tttccattcg acccccatta tgtggaagtc
     1861 ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc
     1921 ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg
     1981 acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg
     2041 ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg
     2101 gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag
     2161 cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc
     2221 tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc
     2281 ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc
     2341 gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt
     2401 gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac
     2461 atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg
     2521 ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa
     2581 agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct gcaagaagac
     2641 aacccggacc tgatcggcag cgagatcgcg cgctggctgt cgacgctcga gatttccggc
     2701 ggcagcagcc atcatcatca tcatcatggg tcagggtaat aatcgaattc aaggcctctc
     2761 gagcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
     2821 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2881 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2941 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     3001 cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt
     3061 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3121 attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg
     3181 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3241 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5221 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5281 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5341 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5401 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5461 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5521 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5581 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5641 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     5701 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
     5761 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
     5821 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
     5881 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
     5941 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
     6001 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
     6061 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     6121 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     6181 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     6241 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     6301 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     6361 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     6421 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     6481 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     6541 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     6601 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     6661 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     6721 taagcaatgc tcgtttaggg aac
//
