LOCUS       sequence-221221-        9597 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab27a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9597)
  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 9597)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-rab27a
FEATURES             Location/Qualifiers
     source          1..9597
                     /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(1421..1441)
                     /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(1449..1465)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1481..1615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1505..1524)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1559..1578
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2287..2303)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2303)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2340..2360)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2340..2358)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2379..2395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2392..2414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2403..2419
                     /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(2427..2433,2434..2451,2452..2457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2472..2493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2610..2627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2781..3369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2861..2880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3540..4331,4332..4400))
                     /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     4163..4182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4401..4505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4668..4689)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5585..5601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5621..5641
                     /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 ggtgtccgat ggggactatg attacctcat
      781 caagttcctc gctctggggg actctggggt gggaaagacc agcttcctat atcagtacac
      841 agacagcaaa ttcaactcca aattcataac cacagtggga atagacttca gagaaaaacg
      901 agtggtgtat aaatccagtg gtcctgatgg cacagcaggc agaggccaga ggatccacat
      961 ccagctttgg gatacggccg gacaggagag gtttcgaagt ttgaccactg ctttcttcag
     1021 agatgcgatg ggctttctgc tgctctttga cctcaccaac gagcagagct tcctgaatgt
     1081 gcgcaactgg atgagtcagc tgcagacaca tgcgtactgt gagaatcctg atatcgtctt
     1141 gtgtggaaat aaatcagacc tggaggagca gcgtgccgtc aaagcggaga acgcaagaga
     1201 actggcagag aaatacggcg tgccatactt cgagacgagc gcggcgaacg gtgagaatgt
     1261 gagtcgtgcg gtggaggttc tgctggatct gatcatgaag cgcatggaga gatgtgtgga
     1321 taaatcctgg atccccgacg ggacagtccg cagcaacggc cacagcaccg ccgacctgac
     1381 agaaccacac gaccccgagc agagcaagtg tgcatgctaa caactttatt atacatagtt
     1441 gataattcac tggccgtcgt tttacggtac catcgatgat gatccagaca tgataagata
     1501 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1561 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1621 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     1681 caagtaaaac ctctacaaat gtggtatggc tgattatgat cctctagatc agatctgcga
     1741 agatacggcc acgggtgctc ttgatcctgt ggctgatttt ggactgtgct gctcgcagct
     1801 gctgatgaat cacatacttc ctccattttc ttccactgat tgactgttat aatttcccta
     1861 atttccaggt caaggtgctg tgcattgtgg taatagatgt gacatgacgt cacttccaaa
     1921 ggaccaatga acatgtctga ccaatttcat ataatgtgaa aacgattttc ataggcagaa
     1981 taaataacat ttaaattaaa ctgggcatca gcgcaattca attggtttgg taatagcaag
     2041 ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt
     2101 aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg
     2161 atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat
     2221 caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc ttcagcaagc
     2281 ccagcagatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     2341 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     2401 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2461 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2521 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2581 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2641 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2701 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3181 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3421 tcacgttaag gcattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3541 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3601 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3661 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3721 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3781 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3841 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3901 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3961 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4021 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4081 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4141 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4201 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4261 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4321 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4381 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4501 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4561 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4621 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4681 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4741 gcgatggccc acgctggcta agaactcatc agcctccccg gtccatctac ccacgtacca
     4801 atgcaccaat tggccacaat gacggctact acatggtgcc attccttcct ctttatagga
     4861 atggagacta cctcctgtcc aacaatgctc ttggatacga gtacgcctac ctgttggacc
     4921 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc
     4981 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag
     5041 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt
     5101 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt
     5161 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact
     5221 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt
     5281 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc
     5341 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc
     5401 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa
     5461 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca
     5521 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccct
     5581 cgagcaggaa acagctatga ccatgattac gccaagctat caactttgta tagaaaagtt
     5641 ggcacgcgca gattccgtgt gggcctggta atatcttacc attttaagct attttaagat
     5701 tagggaaaaa aattaaataa aactacttta aaattacttt ccttaaaaag caacatattt
     5761 atttagtttt tgtttaactg gctcaagatt gtaatatttt acttttacat gtttttttaa
     5821 gtacctaaat gtgttcaaaa gtaaagtatg ttattgtttt ttttttttaa tgttagctaa
     5881 gctttaaatt gtaatatatt acttttaaaa gtgacttttt aaaaatgtaa ataaaaaagt
     5941 aactataaat gtgtaatgca tgttatttct gtgggttttt aattttcatc tcaactgggc
     6001 ttacaatttt aacataataa aaatcaaatc aaatcaaatc aggcccttat ttgtcacata
     6061 cactcttgta tactgaaata aactaaatgt tgagaaaaat ttaaatacac caaaataacg
     6121 catacctttt ttgtaattac ctaagtaaaa catttagttc cttttttaag tgactcaaga
     6181 ttgtaatcta ttaattgtaa aagataaaac tagtgatcaa tctgacgtgc ttctatgcag
     6241 cctgtgagat taattgtgat gtttaattat taaacaattt aaatggatca aacatttcac
     6301 atgtgcttat tgtgtccaac aaaaccagca gaagcactta aaaatattgc atgacaacat
     6361 gagtttttaa tgtaaattat atatacttcc taaacgaagt taatcatatt tctaaaataa
     6421 taatgtcatt gaaatattta aataaatgat aagctttagt tgattcaaca taaaattgaa
     6481 agcaataagg actttttttt tttacagcgg tgaagcaaaa gcatgtaata atcatataat
     6541 ataataaata aataaataaa taatggctct tggtgcaaat atgaacactt taaataaata
     6601 tagaataaat atgatcaata agtcataacg atgtgtttta tgttgattat aatttaaaca
     6661 tgtttctaac gcaagtttta aaaactgttt taaagtatta ttttaagtta taagatcaat
     6721 ttgactcaac attttgtagt gaaaatatgt tgtgtaaaat taatataata tacactataa
     6781 aaatgcaggg ttccacacaa tacattaatg ttgtcccaac acaaatcgat taagttaaca
     6841 aatatatgtg gattgaacat aaaaaattaa gttgtcccaa tgaaatctca agaattgtgt
     6901 tgtttcagct cattttaaat aagtagtttt aacaagtaaa aaataacata tttttgagtg
     6961 tgtatatata tatattacaa aatataataa tataaaatgt aaaagatata aaccatgcta
     7021 catatattta agagctattt tgcacattac ttttttgcaa gccattttag tgtttttttt
     7081 acaaatatat attagctaaa tatacaatat cttaaatata caattaatca gacatgcaat
     7141 taatgcattt aacctacata aaaatacatg atcaacatat gttttatgct gattacaact
     7201 aaatcaaacg taatgaaatt aatttataaa agcgatttaa atatgtcaat attttaagtt
     7261 aaaatcactt gaaacctact ataaatgaac ttaaattaaa aaaatctcct ttaattccaa
     7321 tgcaacagtg ttgttttttc agctgaggac ccccggtgca gatggttggc ccccggcccg
     7381 ctattctaag gaagcctttc ttaattttag ggcggcgctg catgcgctca gatcagattc
     7441 acgggtggag agggttattt tctctttttt attttttctt gcctgctcat atttgggcct
     7501 gtgtgaaccc caggctccgc ctcggccccc aggagccgct cttatttaaa ggggatcctc
     7561 gccttcagtc ttcgtcccag tggaagcatc agtcttgtga taacaacttg gtgagtccta
     7621 cagaccagcg ctcgactcta ggctttaaat gcattgattt ctctggatta gctaccggga
     7681 gagttttaac tttgcttatt agtcttgtgc tcgaactgct aggtttaaag agtgttttct
     7741 ctaatgcatg catgaagcga cagctgaaaa ccagaaatag agcgtttgtc ctcctgttaa
     7801 actctagcct ttctttggca tttgtcagtc cgagtcttca gctgccggca ggagattggg
     7861 tttcagtaac acttgctgag cacaaactac acgtgctttc ttcagattta attcattatt
     7921 aatcattcta gatttaaatg atgttcttac tcattgctga ttattttagc cttgcggagc
     7981 agattaaatg agcacgtgtt tcattaagat tgatttaaat ataattaata tttcaccact
     8041 gtagttttaa aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta
     8101 gattactatt ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt
     8161 gattttatta gattctagtt ttaaaatcat ctcttagcct taatgagcag actgaatgaa
     8221 catgtttcat taagattaac ttcattataa tgaattattt aatcattcta gtgggttata
     8281 aatcagctct atttaaatta attgtcgtgt ttatttctat ttaatttttg attatttttt
     8341 cagggtcgtt tttatgtgat tttttcccgt ttctaatttt aaactaattg tttgattttt
     8401 atatgattcg actcttttta aatgaattgt tttgcttttt ttattgagca tatatatatt
     8461 atacaatttc aatcaacaat tgttgtgctt tttaaatgtt gtttctattt catttctagg
     8521 gtcgatttta ttcgattcta gttttaaaat cgtgattttt aacttctaac cttactgagc
     8581 agactgaatg aacatgtttc atcaagatta acttcattat agttaattat tgaatcgtgc
     8641 tagtggttta taaatcagct ctatttaaat taatcatcat cattatttct atttaatttt
     8701 tattatttct ttaggattat ttttatgtga tttttttcac gttctagttt taaactcaat
     8761 gctcattact gctcaccctt gctgagcagt ctttcgttaa gattaaatta attaataacc
     8821 attctagttt taaaacagtg tcttaaggca tagctgatat tttacaagtt tactatttaa
     8881 tatatctcct ttcttttttt agggtcgatt tcatctgatt tgattaattt ttattatctt
     8941 cactaagctt tttgttctag gttttattct cagtattatt actgttatat ttgaactgaa
     9001 tttctcctca tagtttattc attttattta tttatttttt gtttattcgc tattttgtat
     9061 tttagttttt aattagaaat tttattagaa tcgttttgaa ctagttttta tgcactctaa
     9121 cattaatatt aaatcataac tattattttt attataattt atttgtgttt atctttttgt
     9181 gtagctttca tgtgttttta ttttagtttt attatgtttg cattgaattt tttggactag
     9241 tttatataac gatatacaga aaagccatga caacaaatgt ttttataata cctattaagt
     9301 ataagctaat cagatttcca ctattctgtt caaagttaac attattagcc attctgatgg
     9361 atcatttgac atttgtcata tgtctttatg aacactgctc tttcagatgc cacttttaaa
     9421 gctgtttttt acagtaaatg aacgtttatt attatgacat tatttcctaa ggcctctaga
     9481 gtacagcagg atgtggcgtg tgcattaccg tgtaattatg ctgactgtaa atgtgcattt
     9541 atagactgtc agcatgtctg actgtgcgct gttgtgattg gtcctgcaca gaccaag
//
