LOCUS       sequence-380452-        4633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of the hydrogen peroxide sensor Hyper7 from 
            the T3 promoter. Parton lab clone KRL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4633)
  AUTHORS   Wu Y, Lim YW, Stroud DA, Martel N, Hall TE, Lo HP, Ferguson C, Ryan 
            MT, McMahon KA, Parton RG
  TITLE     Caveolae sense oxidative stress through membrane lipid peroxidation 
            and cytosolic release of CAVIN1 to regulate NRF2.
  JOURNAL   Dev Cell. 2023 Mar 13;58(5):376-397.e4. doi: 
            10.1016/j.devcel.2023.02.004. Epub 2023 Feb 28.
  PUBMED    36858041
REFERENCE   2  (bases 1 to 4633)
  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.devcel.2023.02.004"; journalName: "Dev Cell"; date: 
            "2023-03-13- 13"; volume: "58"; issue: "5"; pages: "376-397.e4"
COMMENT     Sequence Label: Hyper7 in pT3TS-Dest
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    93..117
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1564..1584)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1609..1734
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1821..1840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1822..1840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1847..1864)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1847..1863)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1856..1878)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2005..2460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2142..2163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2354..2373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2487..2591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2592..2660,2661..3452)
                     /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(2810..2829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3623..4211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4112..4131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4365..4382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4499..4520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4535..4540,4541..4558,4559..4565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4573..4589
                     /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     4578..4600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4597..4613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4597..4613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctcaagttt gtacaaaaaa gcaggctgcc
      121 accatgcatt tggcgaatga agaacaaaat gagctggagg gtgcgtttaa gttgggcctc
      181 atcttcacgg tggcacccta ccttcttcct aaacttatcg tatctctgcg gaggaccgcg
      241 cctaagatgc ccttgatgct cgaggagaac tacacacata cgctcacaga gtcactcaag
      301 aggggagacg tcgacgcaat cattgtcgca gaaccgttcc aagagccggg tatcgtgacg
      361 gagcctctct acgatgagcc gttcttcgtg atagtcccga agggtcacag tttcgaggag
      421 cttgatgctg tttcccctcg gatgcttggt gaagagcaag ttctcctcct tacagagggg
      481 aattgtatga gggatcaagt actgagcagc gcgggcttta atagcaacaa tgtatatata
      541 atggcagaca aacagaaaaa tggcataaag gcaaatttta agattcgtca taatgtagaa
      601 gacggtagcg ttcaactcgc cgatcactac cagcaaaaca ctccaatcgg tgacgggcct
      661 gtactcctcc cggataacca ttacctctca ttccagagtg tactcagtaa agatcctaac
      721 gagaaaagag atcatatggt cttgttggag tttgttacag cagcggggat caccctgggc
      781 atggatgagc tttataacgt agatggcggt tccggaggaa cgggttctaa gggtgaagaa
      841 ttgtttaccg gagtagttcc tatcttggtc gaactggatg gggacgttaa cggccataaa
      901 ttttcagttt ctggggaagg cgagggagat gcaacttatg gcaagctgac cctcaaactc
      961 atctgcacga cgggaaagct cccagttcct tggcctacgc tggtgaccac gctgtcttat
     1021 ggcttgaagt gttttgcgcg ttacccagac cacatgaagc agcatgactt ttttaagtct
     1081 gccatgcccg aggggtacgt tcaggagaga acaattttct tcaaagacga cgggaactac
     1141 aaaacaagag ccgaagtgaa atttgaaggg gataccttgg tgaatcgcat cgagctgaag
     1201 ggaatcggat tcaaagaaga tggcaacatc cttgggcaca aactcgagta taacggcccg
     1261 agttgtagtg agcttgcggc caagcagcgt atccagggtc ttacaaatac attgcaagga
     1321 tcttctatca acacaattcg gcacatggtc gccagtgggc tcgcaatctc tgtacttccg
     1381 gccaccgcgt tgacggagaa cgatcatatg ctgttctcca tcattccctt tgagggaacg
     1441 ccgccatcca gaagggtggt tctcgcttat cgaaggaact ttgttcggcc aaaggccttg
     1501 tctgctatga aggccgctat tatgcaaagc cagttgcatg gggtgagctt tatatgcgat
     1561 tgacaacttt attatacata gttgctcgag cggatctggt taccactaaa ccagcctcaa
     1621 gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt
     1681 cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattcta
     1741 aaaaaaaaaa aaaaaaaaaa aaaaaaaanc cccccccccc ccctgcaggg ccaagtcggc
     1801 ctctagaggt acccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     1861 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1921 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1981 tgcgcagcct gaatggcgaa tggcaaattg taagcgttaa tattttgtta aaattcgcgt
     2041 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2101 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2161 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2221 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2281 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2341 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2401 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2521 attcaaatat gtatctgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2701 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2761 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2881 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3001 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3121 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3181 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3421 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4321 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4381 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     4441 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     4501 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     4561 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     4621 acgccaagct cga
//
