LOCUS       sequence-224792-        9548 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab9b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9548)
  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 9548)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-rab9b
FEATURES             Location/Qualifiers
     source          1..9548
                     /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(1372..1392)
                     /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(1400..1416)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1432..1566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1456..1475)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1510..1529
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2238..2254)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2238..2254)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2291..2311)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2291..2309)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2330..2346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2343..2365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2354..2370
                     /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(2378..2384,2385..2402,2403..2408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2423..2444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2561..2578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2812..2831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3491..4282,4283..4351))
                     /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     4114..4133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4352..4456)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4619..4640)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5536..5552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5572..5592
                     /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 ggtgagtggg aagagcctct tgctgaaggt
      781 cattcttctg ggggatggcg gcgtgggcaa aagctccctc atgaaccgat atgtgacgga
      841 ccgcttcgac tcgcagtctt tccataccat cggtgtcgaa ttccttaacc gcgacctcga
      901 aatcgatggt cggctggtca ccctacagat ctgggacacg gcaggccagg agcgattcaa
      961 gagcctgcgc acaccttttt accgtggtgc agattgctgt ctgctcacgt tcgcagtgga
     1021 tgacctgcag agcttccaga atctgggctg ctggaaaaag gagttcatgt attattcaga
     1081 tgtacgagac ccggagcgct tcccctttgt agtgctggga aacaaggtag acaaggaaga
     1141 gcgtgaggtc ggtgaggatg aggccagggc ctggtgtgaa gaaaatggct gctgtccgta
     1201 ctttgaaacc agtgcaaagg acgacacaaa tgtgggggct gctttcgaag ccgctgttcg
     1261 ggaggttctg gctagcgagg accccattga ccacaccttg ctgagcagtt ccatcgatct
     1321 ccatggaaac cgtaaagtga cccgctcgtc ctgctgttga ggtgcctcta acaactttat
     1381 tatacatagt tgataattca ctggccgtcg ttttacggta ccatcgatga tgatccagac
     1441 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1501 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1561 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1621 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagat
     1681 cagatctgcg aagatacggc cacgggtgct cttgatcctg tggctgattt tggactgtgc
     1741 tgctcgcagc tgctgatgaa tcacatactt cctccatttt cttccactga ttgactgtta
     1801 taatttccct aatttccagg tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg
     1861 tcacttccaa aggaccaatg aacatgtctg accaatttca tataatgtga aaacgatttt
     1921 cataggcaga ataaataaca tttaaattaa actgggcatc agcgcaattc aattggtttg
     1981 gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt tttgactgta
     2041 aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat taagtaaaag
     2101 taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa ataatactta
     2161 agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt gaccccctac
     2221 cttcagcaag cccagcagat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     2281 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2461 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2521 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa ggcattttgg tcatgagatt atcaaaaagg atcttcacct
     3421 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3481 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3541 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3601 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3661 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3721 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3781 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3841 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3901 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3961 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4021 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4081 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4141 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4201 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4261 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4321 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4501 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4561 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4621 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4681 cgtctatcag ggcgatggcc cacgctggct aagaactcat cagcctcccc ggtccatcta
     4741 cccacgtacc aatgcaccaa ttggccacaa tgacggctac tacatggtgc cattccttcc
     4801 tctttatagg aatggagact acctcctgtc caacaatgct cttggatacg agtacgccta
     4861 cctgttggac ccaggtcatt gcacaacacc agaaatgccc tctgatctgc aaaagacgtg
     4921 aatatctgtt cagacaccca tatccactct gttccacaca ggtcagaggt ttgtccagga
     4981 gttcttgaca gaggtgtaaa aagtactcaa aaattttact caagtgaaag tacaagtact
     5041 tagggaaaat tttactcaat taaaagtaaa agtatctggc tagaatctta cttgagtaaa
     5101 agtaaaaaag tactccatta aaattgtact tgagtattaa ggaagtaaaa gtaaaagcaa
     5161 gaaagaaaac tagagattct tgtttaagct tttaatctca aaaaacatta aatgaaatgc
     5221 atacaaggtt ttatcctgct ttagaactgt ttgtatttaa ttatcaaact ataagacaga
     5281 caatctaatg ccagtacacg ctactcaaag ttgtaaaacc tcagatttaa cttcagtaga
     5341 agctgattct caaaattgtt agtgtcaagc ctagctcttt tggggctgaa aagcaatcct
     5401 gcagtgctga aaagcctctc acaggcagcc gatgcgggaa gaggtgtatt agtcttgata
     5461 gagaggctgc aaatagcagg aaacgtgagc agagactccc tggtgtctga aacacaggcc
     5521 agatgggccc tcgagcagga aacagctatg accatgatta cgccaagcta tcaactttgt
     5581 atagaaaagt tggcacgcgc agattccgtg tgggcctggt aatatcttac cattttaagc
     5641 tattttaaga ttagggaaaa aaattaaata aaactacttt aaaattactt tccttaaaaa
     5701 gcaacatatt tatttagttt ttgtttaact ggctcaagat tgtaatattt tacttttaca
     5761 tgttttttta agtacctaaa tgtgttcaaa agtaaagtat gttattgttt ttttttttta
     5821 atgttagcta agctttaaat tgtaatatat tacttttaaa agtgactttt taaaaatgta
     5881 aataaaaaag taactataaa tgtgtaatgc atgttatttc tgtgggtttt taattttcat
     5941 ctcaactggg cttacaattt taacataata aaaatcaaat caaatcaaat caggccctta
     6001 tttgtcacat acactcttgt atactgaaat aaactaaatg ttgagaaaaa tttaaataca
     6061 ccaaaataac gcataccttt tttgtaatta cctaagtaaa acatttagtt ccttttttaa
     6121 gtgactcaag attgtaatct attaattgta aaagataaaa ctagtgatca atctgacgtg
     6181 cttctatgca gcctgtgaga ttaattgtga tgtttaatta ttaaacaatt taaatggatc
     6241 aaacatttca catgtgctta ttgtgtccaa caaaaccagc agaagcactt aaaaatattg
     6301 catgacaaca tgagttttta atgtaaatta tatatacttc ctaaacgaag ttaatcatat
     6361 ttctaaaata ataatgtcat tgaaatattt aaataaatga taagctttag ttgattcaac
     6421 ataaaattga aagcaataag gacttttttt ttttacagcg gtgaagcaaa agcatgtaat
     6481 aatcatataa tataataaat aaataaataa ataatggctc ttggtgcaaa tatgaacact
     6541 ttaaataaat atagaataaa tatgatcaat aagtcataac gatgtgtttt atgttgatta
     6601 taatttaaac atgtttctaa cgcaagtttt aaaaactgtt ttaaagtatt attttaagtt
     6661 ataagatcaa tttgactcaa cattttgtag tgaaaatatg ttgtgtaaaa ttaatataat
     6721 atacactata aaaatgcagg gttccacaca atacattaat gttgtcccaa cacaaatcga
     6781 ttaagttaac aaatatatgt ggattgaaca taaaaaatta agttgtccca atgaaatctc
     6841 aagaattgtg ttgtttcagc tcattttaaa taagtagttt taacaagtaa aaaataacat
     6901 atttttgagt gtgtatatat atatattaca aaatataata atataaaatg taaaagatat
     6961 aaaccatgct acatatattt aagagctatt ttgcacatta cttttttgca agccatttta
     7021 gtgttttttt tacaaatata tattagctaa atatacaata tcttaaatat acaattaatc
     7081 agacatgcaa ttaatgcatt taacctacat aaaaatacat gatcaacata tgttttatgc
     7141 tgattacaac taaatcaaac gtaatgaaat taatttataa aagcgattta aatatgtcaa
     7201 tattttaagt taaaatcact tgaaacctac tataaatgaa cttaaattaa aaaaatctcc
     7261 tttaattcca atgcaacagt gttgtttttt cagctgagga cccccggtgc agatggttgg
     7321 cccccggccc gctattctaa ggaagccttt cttaatttta gggcggcgct gcatgcgctc
     7381 agatcagatt cacgggtgga gagggttatt ttctcttttt tattttttct tgcctgctca
     7441 tatttgggcc tgtgtgaacc ccaggctccg cctcggcccc caggagccgc tcttatttaa
     7501 aggggatcct cgccttcagt cttcgtccca gtggaagcat cagtcttgtg ataacaactt
     7561 ggtgagtcct acagaccagc gctcgactct aggctttaaa tgcattgatt tctctggatt
     7621 agctaccggg agagttttaa ctttgcttat tagtcttgtg ctcgaactgc taggtttaaa
     7681 gagtgttttc tctaatgcat gcatgaagcg acagctgaaa accagaaata gagcgtttgt
     7741 cctcctgtta aactctagcc tttctttggc atttgtcagt ccgagtcttc agctgccggc
     7801 aggagattgg gtttcagtaa cacttgctga gcacaaacta cacgtgcttt cttcagattt
     7861 aattcattat taatcattct agatttaaat gatgttctta ctcattgctg attattttag
     7921 ccttgcggag cagattaaat gagcacgtgt ttcattaaga ttgatttaaa tataattaat
     7981 atttcaccac tgtagtttta aaattgcctt ctaagtcatg gttgattttt aataggtatc
     8041 ggctctaatt agattactat tttggttttt aaatgttatt tctatttcat ttgtaggctc
     8101 gtttttaatg tgattttatt agattctagt tttaaaatca tctcttagcc ttaatgagca
     8161 gactgaatga acatgtttca ttaagattaa cttcattata atgaattatt taatcattct
     8221 agtgggttat aaatcagctc tatttaaatt aattgtcgtg tttatttcta tttaattttt
     8281 gattattttt tcagggtcgt ttttatgtga ttttttcccg tttctaattt taaactaatt
     8341 gtttgatttt tatatgattc gactcttttt aaatgaattg ttttgctttt tttattgagc
     8401 atatatatat tatacaattt caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt
     8461 tcatttctag ggtcgatttt attcgattct agttttaaaa tcgtgatttt taacttctaa
     8521 ccttactgag cagactgaat gaacatgttt catcaagatt aacttcatta tagttaatta
     8581 ttgaatcgtg ctagtggttt ataaatcagc tctatttaaa ttaatcatca tcattatttc
     8641 tatttaattt ttattatttc tttaggatta tttttatgtg atttttttca cgttctagtt
     8701 ttaaactcaa tgctcattac tgctcaccct tgctgagcag tctttcgtta agattaaatt
     8761 aattaataac cattctagtt ttaaaacagt gtcttaaggc atagctgata ttttacaagt
     8821 ttactattta atatatctcc tttctttttt tagggtcgat ttcatctgat ttgattaatt
     8881 tttattatct tcactaagct ttttgttcta ggttttattc tcagtattat tactgttata
     8941 tttgaactga atttctcctc atagtttatt cattttattt atttattttt tgtttattcg
     9001 ctattttgta ttttagtttt taattagaaa ttttattaga atcgttttga actagttttt
     9061 atgcactcta acattaatat taaatcataa ctattatttt tattataatt tatttgtgtt
     9121 tatctttttg tgtagctttc atgtgttttt attttagttt tattatgttt gcattgaatt
     9181 ttttggacta gtttatataa cgatatacag aaaagccatg acaacaaatg tttttataat
     9241 acctattaag tataagctaa tcagatttcc actattctgt tcaaagttaa cattattagc
     9301 cattctgatg gatcatttga catttgtcat atgtctttat gaacactgct ctttcagatg
     9361 ccacttttaa agctgttttt tacagtaaat gaacgtttat tattatgaca ttatttccta
     9421 aggcctctag agtacagcag gatgtggcgt gtgcattacc gtgtaattat gctgactgta
     9481 aatgtgcatt tatagactgt cagcatgtct gactgtgcgc tgttgtgatt ggtcctgcac
     9541 agaccaag
//
