LOCUS       sequence-264830-        6137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express WT Zebrafish Halo-Tev-HSPB7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6137)
  AUTHORS   Surya SL, Long MJC, Urul DA, Zhao Y, Mercer EJ, EIsaid IM, Evans T, 
            Aye Y
  TITLE     Cardiovascular Small Heat Shock Protein HSPB7 Is a Kinetically 
            Privileged Reactive Electrophilic Species (RES) Sensor.
  JOURNAL   ACS Chem Biol. 2018 Feb 8. doi: 10.1021/acschembio.7b00925.
  PUBMED    29397684
REFERENCE   2  (bases 1 to 6137)
  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"; journalName: "ACS Chem 
            Biol. 2018 Feb 8. doi: 10.1021/acschembio.7b00925."
COMMENT     Sequence Label: pCS2 Halo-Tev- HSPB7 (zebrafish) WT
FEATURES             Location/Qualifiers
     source          1..6137
                     /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             267..1157
                     /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             1158..1178
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2189..2323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2267..2286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2436..2456)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2436..2454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2475..2491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2475..2491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2488..2510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2499..2515
                     /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(2523..2529,2530..2547,2548..2553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2568..2589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2706..2723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2877..3465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2957..2976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3636..4427,4428..4496))
                     /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     4259..4278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4497..4601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4627..5082)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4764..4785)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4976..4995
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5153..6137
                     /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 atcccatcga ttcgaattca aggcctctcg agcactgttg
      121 gcctactgga caagcaggca gacccagctc atataaacac tgaaaaagct gctgagtgga
      181 ttcatggtgt tttgaataag aacttgatca ccggttcaat ttgaagttct gactggatat
      241 cgatttgaga aagcatctca acactaatgg cagaaatcgg tactggcttt ccattcgacc
      301 cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg
      361 gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca
      421 tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat
      481 ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca
      541 tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg
      601 gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca
      661 tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct
      721 tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta
      781 cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc
      841 cgttcctgaa ccctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg
      901 ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt
      961 cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag
     1021 ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga
     1081 atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcga
     1141 cgctcgagat ttccggcgaa aacttgtatt tccagggctc aatgagcgcg agcaattctt
     1201 ctgcctatcg atcggagcgt agttaccatc agacgtcatc ctcctcatct tcatcctcta
     1261 cctctgcaaa cccatacatg gagaagagcc gaggactctt cgctgacgac tttggatctt
     1321 tcatgtgtcc gaaggatgct ttgggatttc caaacagaac aggcactgtt ggaaacatca
     1381 agactcttgg agacacatac cagttcacag tggacgtgca agacttctcg cctgaggatg
     1441 tgatcgtcac cacctcaaac aatcagattg aagttcatgc tgaaaagctt gctagcgatg
     1501 ggacagtgat gaacactttc acccataaat gccggctacc agaagatgtg gaccccactt
     1561 cagtgaagtc cagtttaggg gccgatggga ctctgactat aaaagcacag agaaacacag
     1621 ctaaactgga gcatgcacag acgttccgca cagagatcaa aatctagacc tgtgccctcc
     1681 tgctgtgtct gtttcactgt gataaatgac cttgaaaata tagctactta gccctgcttt
     1741 gtcattcctc catgaccttc tctcatttct gctcattaga cttggatgga atttaatcct
     1801 gtcacatgta ttcaaaacct ttaagatgaa tgcactgcta acgtcagctc tctgctaatg
     1861 acatatcagc ttacaggaca aagtgggctc caaaattctg gaagtacatt taatccaaaa
     1921 tataattgaa acattgaaag atatcagaat atatatggct atacagtaga tatagtactt
     1981 tagtacttta aattacattt agttcaccca aacatgtaaa tcctgccatt atttactcat
     2041 ttccacttgt tccaaaccca attaagttcc gttgaacaca aaataaaatg tgcaatgatg
     2101 gaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa ggccacatgt gctcgagcct
     2161 ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     2221 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2281 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2341 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     2401 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     2461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2521 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3301 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4621 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4681 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4741 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4801 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4861 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4921 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4981 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5041 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     5101 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaccg
     5161 ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg
     5221 ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag
     5281 ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc tggcactgtg
     5341 ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt
     5401 gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg
     5461 ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc tattggccag
     5521 gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg
     5581 acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc
     5641 gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg
     5701 tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa
     5761 atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat
     5821 gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt
     5881 aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact
     5941 attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg
     6001 cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa
     6061 tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca
     6121 atgctcgttt agggaac
//
