LOCUS       sequence-221223-        9567 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to rab2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9567)
  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 9567)
  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-rab2a
FEATURES             Location/Qualifiers
     source          1..9567
                     /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(1391..1411)
                     /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(1419..1435)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1451..1585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1475..1494)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1529..1548
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2257..2273)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2257..2273)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2310..2330)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2310..2328)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2349..2365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2362..2384)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2373..2389
                     /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(2397..2403,2404..2421,2422..2427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2442..2463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2580..2597)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2751..3339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2831..2850)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3510..4301,4302..4370))
                     /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     4133..4152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4371..4475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4638..4659)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5555..5571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5591..5611
                     /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 ggtggcgtac gcgtatctct tcaagtacat
      781 catcatcgga gacacgggtg tgggaaagtc gtgcctatta ttacagttca cagataagcg
      841 ctttcagccg gtgcacgatc tcactattgg tgttgaattt ggagcgcgaa tgatcactat
      901 agatggcaaa cagataaaac ttcagatctg ggatacggct ggacaggagt ccttccgctc
      961 aatcaccagg tcttactaca gaggtgcagc tggagctctg ctagtttatg acatcacaag
     1021 aagagacact tttaatcact tgacaacctg gttagaagat gctcgtcaac attccaactc
     1081 taacatggtc atcatgctca tcggaaataa aagtgatttg gagtcacgga gagaggtgaa
     1141 gaaagaggaa ggagaggcct ttgccagaga acatggcttg atcttcatgg agacttctgc
     1201 aaagaccgca tcaaacgtag aagaggcatt tatcaacaca gctaaggaga tttatgaaaa
     1261 gatccaggag ggagtctttg atataaacaa cgaggctaac ggcattaaga ttggacccca
     1321 gcatgctgcc accaactcca cgatgggcgg cagtcaggga ggacagcaag ccggaggggg
     1381 ctgttgctga caactttatt atacatagtt gataattcac tggccgtcgt tttacggtac
     1441 catcgatgat gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1501 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1561 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1621 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     1681 tgattatgat cctctagatc agatctgcga agatacggcc acgggtgctc ttgatcctgt
     1741 ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc
     1801 ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg
     1861 taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat
     1921 ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca
     1981 gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa
     2041 taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa
     2101 aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa
     2161 tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct
     2221 ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct agagcggccg
     2281 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2341 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2401 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2461 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2521 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2581 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2641 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2701 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2761 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2821 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2881 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2941 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3001 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3061 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3121 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3181 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3241 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3301 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3361 acggggtctg acgctcagtg gaacgaaaac tcacgttaag gcattttggt catgagatta
     3421 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3481 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3541 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3601 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3661 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3721 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3781 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3841 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3901 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3961 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4021 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4081 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4141 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4201 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4261 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4321 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4381 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4441 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4501 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4561 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4621 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4681 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc acgctggcta agaactcatc
     4741 agcctccccg gtccatctac ccacgtacca atgcaccaat tggccacaat gacggctact
     4801 acatggtgcc attccttcct ctttatagga atggagacta cctcctgtcc aacaatgctc
     4861 ttggatacga gtacgcctac ctgttggacc caggtcattg cacaacacca gaaatgccct
     4921 ctgatctgca aaagacgtga atatctgttc agacacccat atccactctg ttccacacag
     4981 gtcagaggtt tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc
     5041 aagtgaaagt acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct
     5101 agaatcttac ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag
     5161 gaagtaaaag taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa
     5221 aaaacattaa atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat
     5281 tatcaaacta taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct
     5341 cagatttaac ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt
     5401 ggggctgaaa agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag
     5461 aggtgtatta gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct
     5521 ggtgtctgaa acacaggcca gatgggccct cgagcaggaa acagctatga ccatgattac
     5581 gccaagctat caactttgta tagaaaagtt ggcacgcgca gattccgtgt gggcctggta
     5641 atatcttacc attttaagct attttaagat tagggaaaaa aattaaataa aactacttta
     5701 aaattacttt ccttaaaaag caacatattt atttagtttt tgtttaactg gctcaagatt
     5761 gtaatatttt acttttacat gtttttttaa gtacctaaat gtgttcaaaa gtaaagtatg
     5821 ttattgtttt ttttttttaa tgttagctaa gctttaaatt gtaatatatt acttttaaaa
     5881 gtgacttttt aaaaatgtaa ataaaaaagt aactataaat gtgtaatgca tgttatttct
     5941 gtgggttttt aattttcatc tcaactgggc ttacaatttt aacataataa aaatcaaatc
     6001 aaatcaaatc aggcccttat ttgtcacata cactcttgta tactgaaata aactaaatgt
     6061 tgagaaaaat ttaaatacac caaaataacg catacctttt ttgtaattac ctaagtaaaa
     6121 catttagttc cttttttaag tgactcaaga ttgtaatcta ttaattgtaa aagataaaac
     6181 tagtgatcaa tctgacgtgc ttctatgcag cctgtgagat taattgtgat gtttaattat
     6241 taaacaattt aaatggatca aacatttcac atgtgcttat tgtgtccaac aaaaccagca
     6301 gaagcactta aaaatattgc atgacaacat gagtttttaa tgtaaattat atatacttcc
     6361 taaacgaagt taatcatatt tctaaaataa taatgtcatt gaaatattta aataaatgat
     6421 aagctttagt tgattcaaca taaaattgaa agcaataagg actttttttt tttacagcgg
     6481 tgaagcaaaa gcatgtaata atcatataat ataataaata aataaataaa taatggctct
     6541 tggtgcaaat atgaacactt taaataaata tagaataaat atgatcaata agtcataacg
     6601 atgtgtttta tgttgattat aatttaaaca tgtttctaac gcaagtttta aaaactgttt
     6661 taaagtatta ttttaagtta taagatcaat ttgactcaac attttgtagt gaaaatatgt
     6721 tgtgtaaaat taatataata tacactataa aaatgcaggg ttccacacaa tacattaatg
     6781 ttgtcccaac acaaatcgat taagttaaca aatatatgtg gattgaacat aaaaaattaa
     6841 gttgtcccaa tgaaatctca agaattgtgt tgtttcagct cattttaaat aagtagtttt
     6901 aacaagtaaa aaataacata tttttgagtg tgtatatata tatattacaa aatataataa
     6961 tataaaatgt aaaagatata aaccatgcta catatattta agagctattt tgcacattac
     7021 ttttttgcaa gccattttag tgtttttttt acaaatatat attagctaaa tatacaatat
     7081 cttaaatata caattaatca gacatgcaat taatgcattt aacctacata aaaatacatg
     7141 atcaacatat gttttatgct gattacaact aaatcaaacg taatgaaatt aatttataaa
     7201 agcgatttaa atatgtcaat attttaagtt aaaatcactt gaaacctact ataaatgaac
     7261 ttaaattaaa aaaatctcct ttaattccaa tgcaacagtg ttgttttttc agctgaggac
     7321 ccccggtgca gatggttggc ccccggcccg ctattctaag gaagcctttc ttaattttag
     7381 ggcggcgctg catgcgctca gatcagattc acgggtggag agggttattt tctctttttt
     7441 attttttctt gcctgctcat atttgggcct gtgtgaaccc caggctccgc ctcggccccc
     7501 aggagccgct cttatttaaa ggggatcctc gccttcagtc ttcgtcccag tggaagcatc
     7561 agtcttgtga taacaacttg gtgagtccta cagaccagcg ctcgactcta ggctttaaat
     7621 gcattgattt ctctggatta gctaccggga gagttttaac tttgcttatt agtcttgtgc
     7681 tcgaactgct aggtttaaag agtgttttct ctaatgcatg catgaagcga cagctgaaaa
     7741 ccagaaatag agcgtttgtc ctcctgttaa actctagcct ttctttggca tttgtcagtc
     7801 cgagtcttca gctgccggca ggagattggg tttcagtaac acttgctgag cacaaactac
     7861 acgtgctttc ttcagattta attcattatt aatcattcta gatttaaatg atgttcttac
     7921 tcattgctga ttattttagc cttgcggagc agattaaatg agcacgtgtt tcattaagat
     7981 tgatttaaat ataattaata tttcaccact gtagttttaa aattgccttc taagtcatgg
     8041 ttgattttta ataggtatcg gctctaatta gattactatt ttggttttta aatgttattt
     8101 ctatttcatt tgtaggctcg tttttaatgt gattttatta gattctagtt ttaaaatcat
     8161 ctcttagcct taatgagcag actgaatgaa catgtttcat taagattaac ttcattataa
     8221 tgaattattt aatcattcta gtgggttata aatcagctct atttaaatta attgtcgtgt
     8281 ttatttctat ttaatttttg attatttttt cagggtcgtt tttatgtgat tttttcccgt
     8341 ttctaatttt aaactaattg tttgattttt atatgattcg actcttttta aatgaattgt
     8401 tttgcttttt ttattgagca tatatatatt atacaatttc aatcaacaat tgttgtgctt
     8461 tttaaatgtt gtttctattt catttctagg gtcgatttta ttcgattcta gttttaaaat
     8521 cgtgattttt aacttctaac cttactgagc agactgaatg aacatgtttc atcaagatta
     8581 acttcattat agttaattat tgaatcgtgc tagtggttta taaatcagct ctatttaaat
     8641 taatcatcat cattatttct atttaatttt tattatttct ttaggattat ttttatgtga
     8701 tttttttcac gttctagttt taaactcaat gctcattact gctcaccctt gctgagcagt
     8761 ctttcgttaa gattaaatta attaataacc attctagttt taaaacagtg tcttaaggca
     8821 tagctgatat tttacaagtt tactatttaa tatatctcct ttcttttttt agggtcgatt
     8881 tcatctgatt tgattaattt ttattatctt cactaagctt tttgttctag gttttattct
     8941 cagtattatt actgttatat ttgaactgaa tttctcctca tagtttattc attttattta
     9001 tttatttttt gtttattcgc tattttgtat tttagttttt aattagaaat tttattagaa
     9061 tcgttttgaa ctagttttta tgcactctaa cattaatatt aaatcataac tattattttt
     9121 attataattt atttgtgttt atctttttgt gtagctttca tgtgttttta ttttagtttt
     9181 attatgtttg cattgaattt tttggactag tttatataac gatatacaga aaagccatga
     9241 caacaaatgt ttttataata cctattaagt ataagctaat cagatttcca ctattctgtt
     9301 caaagttaac attattagcc attctgatgg atcatttgac atttgtcata tgtctttatg
     9361 aacactgctc tttcagatgc cacttttaaa gctgtttttt acagtaaatg aacgtttatt
     9421 attatgacat tatttcctaa ggcctctaga gtacagcagg atgtggcgtg tgcattaccg
     9481 tgtaattatg ctgactgtaa atgtgcattt atagactgtc agcatgtctg actgtgcgct
     9541 gttgtgattg gtcctgcaca gaccaag
//
