LOCUS       sequence-380104-        5160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of mKate2 tagged zebrafish cavin1a from the 
            T3 promoter. The red fluorescent tag is at the N-terminus. Parton 
            lab clone LAI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5160)
  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 5160)
  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: mKate2-DrCavin1a in pT3TS-Dest
FEATURES             Location/Qualifiers
     source          1..5160
                     /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"
     CDS             124..819
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     protein_bind    complement(821..845)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2092..2112)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           2137..2262
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2348..2367)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2349..2367)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2374..2391)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2374..2390)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2383..2405)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2532..2987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2669..2690)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2881..2900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3014..3118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3119..3187,3188..3979)
                     /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(3337..3356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4150..4738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4639..4658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4892..4909
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5026..5047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5062..5067,5068..5085,5086..5092)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5100..5116
                     /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     5105..5127
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5124..5140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5124..5140
                     /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 accatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
      181 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
      241 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
      301 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
      361 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
      421 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
      481 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
      541 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
      601 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
      661 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
      721 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
      781 gccagatact gcgacctccc tagcaaactg gggcacagag acccagcttt cttgtacaaa
      841 gtggtggctg atacagactt taagctggaa agggctattt tcgctgaggt aagcgatgat
      901 gacgaagagg ttgccctggt cacatctgcc gcgaaaccag ccaccagtct taaagaattt
      961 aagtcacggc gaggggatga tgaagatgac gacgatgata gcgagaaaga agttgatttg
     1021 gtattgggac cggggctgga tgggggggac gcagagaagt ctgacgccac tggcatactt
     1081 gtgttggcct tgctggacaa gatcattggt gtagtagaca aaatacaaca aacacaaact
     1141 tcattggaag caaaacagga agtgatggaa aagaatatga gttcaataca gacagaattg
     1201 gcaaaacttg ccaagtcaca cgtcggtacg gcgggtaccg tgaataagct cttggacaaa
     1261 gttaggaaag ttaatgtcaa tgtcaagagt gtgagggccg acttggagaa gcaggtgggg
     1321 cagatcaaaa aattggagaa caatgaacaa gaacttctga aacgcaaaaa cataaaagtg
     1381 ttgataatcc ccgaaaaagg aaagcctgta aaggtgtcaa agaaacctgc cggtgaagag
     1441 ggagagcagg ttctgtccga gggtggtgag gaagagcatg cttctgaggg agaggaggtc
     1501 gaggtagaag agaccattga agaaagtaga gctgaaagaa ttaaacgatc cggtctccag
     1561 aaggtcgatg aattgaaaaa agcattctcc aaggagcaga tgcaaaagac caggcagaaa
     1621 actaaggaaa atctcgagaa gacaaggatg cggactaaag agaatttgga gaagacacgg
     1681 caacgcacgc gcgagaatct cgaaaagact aagaagtccc ttggtaaaaa aatgggcaaa
     1741 ttcggcacca agcttacccc gaacactgag agaagggaga agataaggtc atcccgtgaa
     1801 aaaatgaaaa aaagccttaa cccggatcac accgtgtatg cccgctcaaa gtctacgact
     1861 tatcgcgtac caccattcac attccatgtc aagaaaattc gtggtggtga agcagagccg
     1921 acaccggagc cagaagagga agatgaagag cagaaggaag aagaggaaga aggcgacctg
     1981 gttgaggagg ccgaactcgt tagccttgat ggaccagaaa tggaggcggc tgatcgatct
     2041 cgcctcgcct ttcaggaaac cgttcgccct agagagaccg ctggcctgta gcaactttat
     2101 tatacatagt tgctcgagcg gatctggtta ccactaaacc agcctcaaga acacccgaat
     2161 ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt
     2221 agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa
     2281 aaaaaaaaaa aaaaaamccc cccccccccc tgcagggcca agtcggcctc tagaggtacc
     2341 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     2401 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2461 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2521 tggcgaatgg caaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2581 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2641 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2701 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2761 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2821 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2881 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2941 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3001 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3061 tctgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3121 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3181 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3241 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3301 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3361 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3421 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3481 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3541 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3601 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3661 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3721 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3781 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3841 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3901 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3961 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4861 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4921 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     4981 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     5041 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5101 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcga
//
