LOCUS       sequence-220543-        9596 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab11bb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9596)
  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 9596)
  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-rab11bb
FEATURES             Location/Qualifiers
     source          1..9596
                     /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(1420..1440)
                     /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(1448..1464)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1480..1614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1504..1523)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1558..1577
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2286..2302)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2286..2302)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2339..2359)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2339..2357)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2378..2394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2391..2413)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2402..2418
                     /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(2426..2432,2433..2450,2451..2456))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2471..2492
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2609..2626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2780..3368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2860..2879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3539..4330,4331..4399))
                     /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     4162..4181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4400..4504)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4667..4688)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5584..5600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5620..5640
                     /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 ggtggccaac agagacgatg aatacgattt
      781 cttgttcaaa gtcgttctca tcggagattc aggcgtcggg aagagtaacc tgctgtcgag
      841 attcacacga aacgagttta acctggagag caagagcacg ataggagtgg agttcgccac
      901 caggagtatt caggtggatg gaaagaccat aaaagcacag atctgggata cggctggaca
      961 ggagcgatac agagccatca cttctgcata ttatcgtgga gcggtcggtg cgctgctggt
     1021 gtacgacatc gccaaacacc ttacctatga gaacgtggag cgctggctga aagagctgcg
     1081 agatcacgct gacaacaaca tcgtcattat gctggttggg aataagagtg acctgcggca
     1141 cctgagggcc gtgcccactg atgaggcccg cgctttcgca gagaagaata acctgtcatt
     1201 catcgagacg tcagctctgg actcaactaa tgttgaagag gcattcaaaa acatcctaac
     1261 agaaatctac cgtatcgtat cacagaagca gatagcggag cggtcggctc atgacgagtc
     1321 tcccggaaat aacgttgtgg acatcagtgt cccgcccacc acagacggac agaagagcaa
     1381 caaactgcag tgctgccaaa acctgtaacc acagcgaccc aactttatta tacatagttg
     1441 ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat gataagatac
     1501 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1561 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1621 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1681 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca gatctgcgaa
     1741 gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg ctcgcagctg
     1801 ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata atttccctaa
     1861 tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc acttccaaag
     1921 gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca taggcagaat
     1981 aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt aatagcaagg
     2041 gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa taaaattgta
     2101 aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta aaagtattga
     2161 tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag tacagtaatc
     2221 aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct tcagcaagcc
     2281 cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     2341 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2401 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2461 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2521 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2581 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2641 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2701 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2761 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2821 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2881 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2941 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3001 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3061 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3121 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3181 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3241 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3301 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3361 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3421 cacgttaagg cattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4501 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     4561 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4621 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4681 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4741 cgatggccca cgctggctaa gaactcatca gcctccccgg tccatctacc cacgtaccaa
     4801 tgcaccaatt ggccacaatg acggctacta catggtgcca ttccttcctc tttataggaa
     4861 tggagactac ctcctgtcca acaatgctct tggatacgag tacgcctacc tgttggaccc
     4921 aggtcattgc acaacaccag aaatgccctc tgatctgcaa aagacgtgaa tatctgttca
     4981 gacacccata tccactctgt tccacacagg tcagaggttt gtccaggagt tcttgacaga
     5041 ggtgtaaaaa gtactcaaaa attttactca agtgaaagta caagtactta gggaaaattt
     5101 tactcaatta aaagtaaaag tatctggcta gaatcttact tgagtaaaag taaaaaagta
     5161 ctccattaaa attgtacttg agtattaagg aagtaaaagt aaaagcaaga aagaaaacta
     5221 gagattcttg tttaagcttt taatctcaaa aaacattaaa tgaaatgcat acaaggtttt
     5281 atcctgcttt agaactgttt gtatttaatt atcaaactat aagacagaca atctaatgcc
     5341 agtacacgct actcaaagtt gtaaaacctc agatttaact tcagtagaag ctgattctca
     5401 aaattgttag tgtcaagcct agctcttttg gggctgaaaa gcaatcctgc agtgctgaaa
     5461 agcctctcac aggcagccga tgcgggaaga ggtgtattag tcttgataga gaggctgcaa
     5521 atagcaggaa acgtgagcag agactccctg gtgtctgaaa cacaggccag atgggccctc
     5581 gagcaggaaa cagctatgac catgattacg ccaagctatc aactttgtat agaaaagttg
     5641 gcacgcgcag attccgtgtg ggcctggtaa tatcttacca ttttaagcta ttttaagatt
     5701 agggaaaaaa attaaataaa actactttaa aattactttc cttaaaaagc aacatattta
     5761 tttagttttt gtttaactgg ctcaagattg taatatttta cttttacatg tttttttaag
     5821 tacctaaatg tgttcaaaag taaagtatgt tattgttttt tttttttaat gttagctaag
     5881 ctttaaattg taatatatta cttttaaaag tgacttttta aaaatgtaaa taaaaaagta
     5941 actataaatg tgtaatgcat gttatttctg tgggttttta attttcatct caactgggct
     6001 tacaatttta acataataaa aatcaaatca aatcaaatca ggcccttatt tgtcacatac
     6061 actcttgtat actgaaataa actaaatgtt gagaaaaatt taaatacacc aaaataacgc
     6121 ataccttttt tgtaattacc taagtaaaac atttagttcc ttttttaagt gactcaagat
     6181 tgtaatctat taattgtaaa agataaaact agtgatcaat ctgacgtgct tctatgcagc
     6241 ctgtgagatt aattgtgatg tttaattatt aaacaattta aatggatcaa acatttcaca
     6301 tgtgcttatt gtgtccaaca aaaccagcag aagcacttaa aaatattgca tgacaacatg
     6361 agtttttaat gtaaattata tatacttcct aaacgaagtt aatcatattt ctaaaataat
     6421 aatgtcattg aaatatttaa ataaatgata agctttagtt gattcaacat aaaattgaaa
     6481 gcaataagga cttttttttt ttacagcggt gaagcaaaag catgtaataa tcatataata
     6541 taataaataa ataaataaat aatggctctt ggtgcaaata tgaacacttt aaataaatat
     6601 agaataaata tgatcaataa gtcataacga tgtgttttat gttgattata atttaaacat
     6661 gtttctaacg caagttttaa aaactgtttt aaagtattat tttaagttat aagatcaatt
     6721 tgactcaaca ttttgtagtg aaaatatgtt gtgtaaaatt aatataatat acactataaa
     6781 aatgcagggt tccacacaat acattaatgt tgtcccaaca caaatcgatt aagttaacaa
     6841 atatatgtgg attgaacata aaaaattaag ttgtcccaat gaaatctcaa gaattgtgtt
     6901 gtttcagctc attttaaata agtagtttta acaagtaaaa aataacatat ttttgagtgt
     6961 gtatatatat atattacaaa atataataat ataaaatgta aaagatataa accatgctac
     7021 atatatttaa gagctatttt gcacattact tttttgcaag ccattttagt gtttttttta
     7081 caaatatata ttagctaaat atacaatatc ttaaatatac aattaatcag acatgcaatt
     7141 aatgcattta acctacataa aaatacatga tcaacatatg ttttatgctg attacaacta
     7201 aatcaaacgt aatgaaatta atttataaaa gcgatttaaa tatgtcaata ttttaagtta
     7261 aaatcacttg aaacctacta taaatgaact taaattaaaa aaatctcctt taattccaat
     7321 gcaacagtgt tgttttttca gctgaggacc cccggtgcag atggttggcc cccggcccgc
     7381 tattctaagg aagcctttct taattttagg gcggcgctgc atgcgctcag atcagattca
     7441 cgggtggaga gggttatttt ctctttttta ttttttcttg cctgctcata tttgggcctg
     7501 tgtgaacccc aggctccgcc tcggccccca ggagccgctc ttatttaaag gggatcctcg
     7561 ccttcagtct tcgtcccagt ggaagcatca gtcttgtgat aacaacttgg tgagtcctac
     7621 agaccagcgc tcgactctag gctttaaatg cattgatttc tctggattag ctaccgggag
     7681 agttttaact ttgcttatta gtcttgtgct cgaactgcta ggtttaaaga gtgttttctc
     7741 taatgcatgc atgaagcgac agctgaaaac cagaaataga gcgtttgtcc tcctgttaaa
     7801 ctctagcctt tctttggcat ttgtcagtcc gagtcttcag ctgccggcag gagattgggt
     7861 ttcagtaaca cttgctgagc acaaactaca cgtgctttct tcagatttaa ttcattatta
     7921 atcattctag atttaaatga tgttcttact cattgctgat tattttagcc ttgcggagca
     7981 gattaaatga gcacgtgttt cattaagatt gatttaaata taattaatat ttcaccactg
     8041 tagttttaaa attgccttct aagtcatggt tgatttttaa taggtatcgg ctctaattag
     8101 attactattt tggtttttaa atgttatttc tatttcattt gtaggctcgt ttttaatgtg
     8161 attttattag attctagttt taaaatcatc tcttagcctt aatgagcaga ctgaatgaac
     8221 atgtttcatt aagattaact tcattataat gaattattta atcattctag tgggttataa
     8281 atcagctcta tttaaattaa ttgtcgtgtt tatttctatt taatttttga ttattttttc
     8341 agggtcgttt ttatgtgatt ttttcccgtt tctaatttta aactaattgt ttgattttta
     8401 tatgattcga ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatta
     8461 tacaatttca atcaacaatt gttgtgcttt ttaaatgttg tttctatttc atttctaggg
     8521 tcgattttat tcgattctag ttttaaaatc gtgattttta acttctaacc ttactgagca
     8581 gactgaatga acatgtttca tcaagattaa cttcattata gttaattatt gaatcgtgct
     8641 agtggtttat aaatcagctc tatttaaatt aatcatcatc attatttcta tttaattttt
     8701 attatttctt taggattatt tttatgtgat ttttttcacg ttctagtttt aaactcaatg
     8761 ctcattactg ctcacccttg ctgagcagtc tttcgttaag attaaattaa ttaataacca
     8821 ttctagtttt aaaacagtgt cttaaggcat agctgatatt ttacaagttt actatttaat
     8881 atatctcctt tcttttttta gggtcgattt catctgattt gattaatttt tattatcttc
     8941 actaagcttt ttgttctagg ttttattctc agtattatta ctgttatatt tgaactgaat
     9001 ttctcctcat agtttattca ttttatttat ttattttttg tttattcgct attttgtatt
     9061 ttagttttta attagaaatt ttattagaat cgttttgaac tagtttttat gcactctaac
     9121 attaatatta aatcataact attattttta ttataattta tttgtgttta tctttttgtg
     9181 tagctttcat gtgtttttat tttagtttta ttatgtttgc attgaatttt ttggactagt
     9241 ttatataacg atatacagaa aagccatgac aacaaatgtt tttataatac ctattaagta
     9301 taagctaatc agatttccac tattctgttc aaagttaaca ttattagcca ttctgatgga
     9361 tcatttgaca tttgtcatat gtctttatga acactgctct ttcagatgcc acttttaaag
     9421 ctgtttttta cagtaaatga acgtttatta ttatgacatt atttcctaag gcctctagag
     9481 tacagcagga tgtggcgtgt gcattaccgt gtaattatgc tgactgtaaa tgtgcattta
     9541 tagactgtca gcatgtctga ctgtgcgctg ttgtgattgg tcctgcacag accaag
//
