LOCUS       sequence-380451-        5278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of mKate2 tagged zebrafish cavin1b from the 
            T3 promoter. The red fluorescent tag is at the N-terminus. Parton 
            lab clone LAC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5278)
  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 5278)
  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-DrCavin1b in pT3TS-Dest
FEATURES             Location/Qualifiers
     source          1..5278
                     /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(2209..2229)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     3'UTR           2254..2379
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2466..2485)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2467..2485)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2492..2509)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2492..2508)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2501..2523)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2650..3105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2787..2808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2999..3018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3132..3236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3237..3305,3306..4097)
                     /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(3455..3474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4268..4856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4757..4776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5010..5027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5144..5165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5180..5185,5186..5203,5204..5210)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5218..5234
                     /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     5223..5245
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5242..5258
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5242..5258
                     /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 gtggtggagg tcattgaaga tagtcgtttg catcttgaag tacccgaact gcggaagtca
      901 ttgacggaga taagcgagga tgaaatcaat ctcccagtct ctgacgatga ggacgaggct
      961 acagctgaaa ataaagccaa agatgcagcg gaaaaggaag aagctggcga gggcggcgag
     1021 atgaacgggg taatggtcct cacattgctt gataaaatta tcggtgcagt ggatcagatt
     1081 caacaaacac aggctggcct tgaagctcgt cagcgagaca tggagcgatc cgtaaccggc
     1141 atccagggag agctgacgaa gcttagtaag tctcatacaa ctactgcgaa cactgtaaac
     1201 aaaatgttgg agaaggtgcg taaagtaagt gtgaacgtga aatccgtccg ggggaacctg
     1261 gaaaaacaag caggccaaat taagcgcctg gagagtaatg agaatgaact cttgaaacga
     1321 cgcaatttta aggtgatgat ataccaggac gaggtcaagg taggatcaaa ggtgaatgtt
     1381 gcaaagagca tgaagtaccc tgagcctgta gagggagagg ccgaaggaga gttgaaagcg
     1441 gcggcggagg agggaggaga ggacaaagaa aaggtgaatc tggacctgtc cagtgatgaa
     1501 gaggaagtcg agatagagga gacaatcgag gaatcccgag cggaacgaat caaacgatct
     1561 agcctgcaga gagtacagaa catcaagact gttttctcta aagggaacat ggagaagaca
     1621 aagcagaaaa cgaaggaaaa cttggagaag acaaaacaga agacgaagga aaacctcgaa
     1681 aaaacgcgga tgcgcaccaa ggagaacctt gagaagaccc gtcagaaaac caaggaaaac
     1741 cttgagaaaa cgaagcaaaa aacgcgggaa aatctggaaa agacaagaca caatattgaa
     1801 aagaagatgg gtaagctggg cactaggatg tcagtcaatc cagagcgtaa ggagaagatg
     1861 aaaacaagcc agaaaaaggt catgaagagt ttcactcccg atcataccat ttacgcgcgc
     1921 tccaagactg cggtgtacaa ggtccctccc ttcacctttc acgtcaagaa gatccgtgag
     1981 ggtgaggtcg agattcaggg gacggaaatg gtggaagtgg taggggacac cgaggaggtt
     2041 gaaaacgcgg taatggaggg ggaggaggat gctgagatgg aaatggaggc gggcgagatg
     2101 gtcgaggaag acgaagacga gactgaggac tcacctgaaa tgcgcgcact tcttgagaag
     2161 ggagatgaat tggtattggt agaggaccat gacagagatt ctgactagca actttattat
     2221 acatagttgc tcgagcggat ctggttacca ctaaaccagc ctcaagaaca cccgaatgga
     2281 gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc
     2341 cattcgtatc tgctcctaat aaaaagaaag tttcttcaca ttctaaaaaa aaaaaaaaaa
     2401 aaaaaaaaaa aaancccccc cccccccctg cagggccaag tcggcctcta gaggtaccca
     2461 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2521 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2581 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2641 gcgaatggca aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2701 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2761 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2821 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     2881 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     2941 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3001 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3061 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3121 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3181 tgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3241 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3301 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3361 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3421 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3481 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3541 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3601 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3661 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3721 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3781 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3841 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3901 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3961 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4021 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4081 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4141 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4201 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4381 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4621 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4921 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4981 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5041 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5101 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5161 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5221 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcga
//
