LOCUS       sequence-383997-        4330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of the redox sensor GLRX-roGFP2 from the T3 
            promoter. Parton lab clone KRK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4330)
  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 4330)
  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: GLRX-roGFP2 in pT3TS-Dest
FEATURES             Location/Qualifiers
     source          1..4330
                     /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"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1261..1281)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           1306..1431
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1518..1537)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1519..1537)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1544..1561)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1544..1560)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1553..1575)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1702..2157)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1839..1860)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2051..2070
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2184..2288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2289..2357,2358..3149)
                     /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(2507..2526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3320..3908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3809..3828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4062..4079
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4196..4217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4232..4237,4238..4255,4256..4262)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4270..4286
                     /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     4275..4297
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4294..4310
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4294..4310
                     /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 accatggccc aggaatttgt taattgcaag atacaacccg gcaaggttgt agtgtttatc
      181 aagccgacct gtccttactg ccgacgtgcg caagaaattc tttcacagct gccaataaag
      241 caggggttgt tggagttcgt tgatattaca gcaactaacc acaccaatga gatccaagac
      301 tatttgcaac aacttaccgg tgcgcgaaca gttccaaggg tctttatcgg aaaggattgt
      361 ataggcggct gctctgacct tgtctctctg caacaatctg gcgagctcct gacgaggttg
      421 aaacaaatag gcgcactgca gacttctgga ggtagtggag ggggtggttc aggcggtggc
      481 ggttctggtg gtggaggtag cgggggcggc ggcagcggag gaggcggatc cgggggagag
      541 tttgtgagta agggtgagga actctttact ggtgttgtcc cgatcctggt ggagctcgat
      601 ggggatgtga acgggcacaa attctccgtc agcggggagg gcgaaggtga cgccacgtac
      661 gggaaattga cgcttaaatt tattagtacc acaggaaagc tgccagtccc gtggccaacc
      721 ctcgtaacga cccttacgta cggagtgcaa tgcttttcca ggtaccctga tcacatgaag
      781 caacacgact tcttcaaatc cgccatgcct gaaggttatg ttcaagaacg gaccatcttt
      841 ttcaaagacg acggcaacta taagacaaga gcagaggtca agtttgaggg tgatacactt
      901 gtcaatagaa tcgagctgaa agggattgac ttcaaggagg atggtaatat cctggggcat
      961 aagctcgagt acaactacaa ctgccataac gtctacatta tggccgataa acagaagaac
     1021 gggattaaag taaatttcaa gattcgtcat aacattgaag acggcagcgt tcaacttgca
     1081 gatcattacc agcaaaacac gcccattgga gacgggccgg ttctgcttcc cgacaaccat
     1141 tacttgagca cgtgtagtgc cctctctaag gacccgaacg agaagcggga tcacatggtt
     1201 cttcttgagt ttgtaacagc agccgggata accctgggta tggatgagtt gtacaaataa
     1261 caactttatt atacatagtt gctcgagcgg atctggttac cactaaacca gcctcaagaa
     1321 cacccgaatg gagtctctaa gctacataat accaacttac actttacaaa atgttgtccc
     1381 ccaaaatgta gccattcgta tctgctccta ataaaaagaa agtttcttca cattctaaaa
     1441 aaaaaaaaaa aaaaaaaaaa aaaaaccccc cccccccccc tgcagggcca agtcggcctc
     1501 tagaggtacc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     1561 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1621 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1681 gcagcctgaa tggcgaatgg caaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1741 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1801 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1861 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1921 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1981 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2041 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2101 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2161 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2221 caaatatgta tctgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2281 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2341 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2401 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2461 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2521 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2581 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2641 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2701 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2761 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2821 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2881 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2941 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3001 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3061 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3121 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3181 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3241 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3301 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3361 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3421 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3481 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3541 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3601 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3661 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3721 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3781 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3841 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3901 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3961 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4021 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4081 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4141 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     4201 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4261 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     4321 ccaagctcga
//
