LOCUS       sequence-224848-        9471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to dominant 
            negative ARF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9471)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 9471)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: actc1b-mKate2-ARF1-DN
FEATURES             Location/Qualifiers
     source          1..9471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             32..727
                     /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(729..753)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1295..1315)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(1323..1339)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1355..1489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1379..1398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1433..1452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1928..2115,2116..2127)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2161..2177)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2161..2177)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2214..2234)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2214..2232)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2253..2269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2266..2288)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2277..2293
                     /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(2301..2307,2308..2325,2326..2331))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2346..2367
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2484..2501)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2655..3243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2735..2754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3414..4205,4206..4274))
                     /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     4037..4056
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4275..4379)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4542..4563)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(4912..4923,4924..5086)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5459..5475
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5495..5515
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 gcaagtttgt acaaaaaagc aggctgccac catggtgagc gagctgatta aggagaacat
       61 gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
      121 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg
      181 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt
      241 catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac
      301 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
      361 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa
      421 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc
      481 cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca
      541 cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
      601 gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac
      661 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg
      721 gcacagagac ccagctttct tgtacaaagt ggtgggaaac atattcgcaa acctctttaa
      781 agtcttcggc aagaaggaga tgagaattct tatggtggga ctcgacgcag cagggaagaa
      841 cacgattctg tataaattaa agttggggga aattgtgacc accatcccaa caatcggttt
      901 taatgttgaa acagtggagt acaagaatat tagttttaca gtttgggacg tcggcggtca
      961 agataagatt cgaccattgt ggcgtcatta tttccagaac acacaaggtc tgatctttgt
     1021 ggttgacagt aatgaccgtg agcgtgtgaa tgaggcgcga gaggagctga tgaggatgtt
     1081 ggctgaggat gagctcagag aagcagttct gctggtgttt gccaacaaac aggacctgcc
     1141 gaacgccatg aacgccgccg aaatcacgga caagctgggt ctgcactcac tccggcacag
     1201 aaactggtac attcaggcca cgtgtgccac aagcggcgac ggcctctatg aaggactcga
     1261 ctggttgtcc aatcagctga aaaaccagaa atgacaactt tattatacat agttgataat
     1321 tcactggccg tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga
     1381 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1441 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1501 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1561 aaacctctac aaatgtggta tggctgatta tgatcctcta gatcagatct gcgaagatac
     1621 ggccacgggt gctcttgatc ctgtggctga ttttggactg tgctgctcgc agctgctgat
     1681 gaatcacata cttcctccat tttcttccac tgattgactg ttataatttc cctaatttcc
     1741 aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg acgtcacttc caaaggacca
     1801 atgaacatgt ctgaccaatt tcatataatg tgaaaacgat tttcataggc agaataaata
     1861 acatttaaat taaactgggc atcagcgcaa ttcaattggt ttggtaatag caagggaaaa
     1921 tagaatgaag tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag
     1981 taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt
     2041 aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta
     2101 aaattactca agtactttac acctctggtt cttgaccccc taccttcagc aagcccagca
     2161 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     2221 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2281 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2341 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2401 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2461 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2521 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2581 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2641 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2701 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2761 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2821 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2881 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2941 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3001 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3061 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3121 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3181 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3241 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3301 taaggcattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3361 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3421 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3481 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3541 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3601 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3661 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3721 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3781 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3841 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3901 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3961 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4021 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4081 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4141 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4201 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4261 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4321 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4381 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4441 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4501 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4561 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4621 gcccacgctg gctaagaact catcagcctc cccggtccat ctacccacgt accaatgcac
     4681 caattggcca caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag
     4741 actacctcct gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc
     4801 attgcacaac accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac
     4861 ccatatccac tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt
     4921 aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc
     4981 aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca
     5041 ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa aactagagat
     5101 tcttgtttaa gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct
     5161 gctttagaac tgtttgtatt taattatcaa actataagac agacaatcta atgccagtac
     5221 acgctactca aagttgtaaa acctcagatt taacttcagt agaagctgat tctcaaaatt
     5281 gttagtgtca agcctagctc ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct
     5341 ctcacaggca gccgatgcgg gaagaggtgt attagtcttg atagagaggc tgcaaatagc
     5401 aggaaacgtg agcagagact ccctggtgtc tgaaacacag gccagatggg ccctcgagca
     5461 ggaaacagct atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttggcacg
     5521 cgcagattcc gtgtgggcct ggtaatatct taccatttta agctatttta agattaggga
     5581 aaaaaattaa ataaaactac tttaaaatta ctttccttaa aaagcaacat atttatttag
     5641 tttttgttta actggctcaa gattgtaata ttttactttt acatgttttt ttaagtacct
     5701 aaatgtgttc aaaagtaaag tatgttattg tttttttttt ttaatgttag ctaagcttta
     5761 aattgtaata tattactttt aaaagtgact ttttaaaaat gtaaataaaa aagtaactat
     5821 aaatgtgtaa tgcatgttat ttctgtgggt ttttaatttt catctcaact gggcttacaa
     5881 ttttaacata ataaaaatca aatcaaatca aatcaggccc ttatttgtca catacactct
     5941 tgtatactga aataaactaa atgttgagaa aaatttaaat acaccaaaat aacgcatacc
     6001 ttttttgtaa ttacctaagt aaaacattta gttccttttt taagtgactc aagattgtaa
     6061 tctattaatt gtaaaagata aaactagtga tcaatctgac gtgcttctat gcagcctgtg
     6121 agattaattg tgatgtttaa ttattaaaca atttaaatgg atcaaacatt tcacatgtgc
     6181 ttattgtgtc caacaaaacc agcagaagca cttaaaaata ttgcatgaca acatgagttt
     6241 ttaatgtaaa ttatatatac ttcctaaacg aagttaatca tatttctaaa ataataatgt
     6301 cattgaaata tttaaataaa tgataagctt tagttgattc aacataaaat tgaaagcaat
     6361 aaggactttt tttttttaca gcggtgaagc aaaagcatgt aataatcata taatataata
     6421 aataaataaa taaataatgg ctcttggtgc aaatatgaac actttaaata aatatagaat
     6481 aaatatgatc aataagtcat aacgatgtgt tttatgttga ttataattta aacatgtttc
     6541 taacgcaagt tttaaaaact gttttaaagt attattttaa gttataagat caatttgact
     6601 caacattttg tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc
     6661 agggttccac acaatacatt aatgttgtcc caacacaaat cgattaagtt aacaaatata
     6721 tgtggattga acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc
     6781 agctcatttt aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata
     6841 tatatatatt acaaaatata ataatataaa atgtaaaaga tataaaccat gctacatata
     6901 tttaagagct attttgcaca ttactttttt gcaagccatt ttagtgtttt ttttacaaat
     6961 atatattagc taaatataca atatcttaaa tatacaatta atcagacatg caattaatgc
     7021 atttaaccta cataaaaata catgatcaac atatgtttta tgctgattac aactaaatca
     7081 aacgtaatga aattaattta taaaagcgat ttaaatatgt caatatttta agttaaaatc
     7141 acttgaaacc tactataaat gaacttaaat taaaaaaatc tcctttaatt ccaatgcaac
     7201 agtgttgttt tttcagctga ggacccccgg tgcagatggt tggcccccgg cccgctattc
     7261 taaggaagcc tttcttaatt ttagggcggc gctgcatgcg ctcagatcag attcacgggt
     7321 ggagagggtt attttctctt ttttattttt tcttgcctgc tcatatttgg gcctgtgtga
     7381 accccaggct ccgcctcggc ccccaggagc cgctcttatt taaaggggat cctcgccttc
     7441 agtcttcgtc ccagtggaag catcagtctt gtgataacaa cttggtgagt cctacagacc
     7501 agcgctcgac tctaggcttt aaatgcattg atttctctgg attagctacc gggagagttt
     7561 taactttgct tattagtctt gtgctcgaac tgctaggttt aaagagtgtt ttctctaatg
     7621 catgcatgaa gcgacagctg aaaaccagaa atagagcgtt tgtcctcctg ttaaactcta
     7681 gcctttcttt ggcatttgtc agtccgagtc ttcagctgcc ggcaggagat tgggtttcag
     7741 taacacttgc tgagcacaaa ctacacgtgc tttcttcaga tttaattcat tattaatcat
     7801 tctagattta aatgatgttc ttactcattg ctgattattt tagccttgcg gagcagatta
     7861 aatgagcacg tgtttcatta agattgattt aaatataatt aatatttcac cactgtagtt
     7921 ttaaaattgc cttctaagtc atggttgatt tttaataggt atcggctcta attagattac
     7981 tattttggtt tttaaatgtt atttctattt catttgtagg ctcgttttta atgtgatttt
     8041 attagattct agttttaaaa tcatctctta gccttaatga gcagactgaa tgaacatgtt
     8101 tcattaagat taacttcatt ataatgaatt atttaatcat tctagtgggt tataaatcag
     8161 ctctatttaa attaattgtc gtgtttattt ctatttaatt tttgattatt ttttcagggt
     8221 cgtttttatg tgattttttc ccgtttctaa ttttaaacta attgtttgat ttttatatga
     8281 ttcgactctt tttaaatgaa ttgttttgct ttttttattg agcatatata tattatacaa
     8341 tttcaatcaa caattgttgt gctttttaaa tgttgtttct atttcatttc tagggtcgat
     8401 tttattcgat tctagtttta aaatcgtgat ttttaacttc taaccttact gagcagactg
     8461 aatgaacatg tttcatcaag attaacttca ttatagttaa ttattgaatc gtgctagtgg
     8521 tttataaatc agctctattt aaattaatca tcatcattat ttctatttaa tttttattat
     8581 ttctttagga ttatttttat gtgatttttt tcacgttcta gttttaaact caatgctcat
     8641 tactgctcac ccttgctgag cagtctttcg ttaagattaa attaattaat aaccattcta
     8701 gttttaaaac agtgtcttaa ggcatagctg atattttaca agtttactat ttaatatatc
     8761 tcctttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa
     8821 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc
     8881 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt
     8941 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa
     9001 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct
     9061 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata
     9121 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc
     9181 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt
     9241 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt
     9301 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag
     9361 caggatgtgg cgtgtgcatt accgtgtaat tatgctgact gtaaatgtgc atttatagac
     9421 tgtcagcatg tctgactgtg cgctgttgtg attggtcctg cacagaccaa g
//
