LOCUS       Exported                9693 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Muscle specific zebrafish expression of mKate2 fused to 3xPHD domain
            from Mouse inhibitor of growth family member 2 (ING2), codon 
            optimised. Binds PtdIns5P.
ACCESSION   .
VERSION     .
KEYWORDS    actc1b-mKate2-ING2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9693)
  TITLE     Parton Lab Plasmids for in vivo cell biology in the zebrafish
REFERENCE   2  (bases 1 to 9693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9693
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    5092..5116
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5117..5126
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5123..5818
                     /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(5820..5844)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6590..6610)
                     /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(6618..6634)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6650..6784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6674..6693)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6728..6747
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7456..7472)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7456..7472)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7509..7529)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7509..7527)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7548..7564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7561..7583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7572..7588
                     /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(7596..7626)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7641..7662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7779..7796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7950..8538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8030..8049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8709..9569)
                     /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     9332..9351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9570..9674)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttggca cgcgcagatt ccgtgtgggc
     1141 ctggtaatat cttaccattt taagctattt taagattagg gaaaaaaatt aaataaaact
     1201 actttaaaat tactttcctt aaaaagcaac atatttattt agtttttgtt taactggctc
     1261 aagattgtaa tattttactt ttacatgttt tttttaagta cctaaatgtg ttcaaaagta
     1321 aagtatgtta ttgttttttt ttttttaatg ttagctaagc tttaaattgt aatatattac
     1381 ttttaaaagt gactttttaa aaatgtaaat aaaaaagtaa ctataaatgt gtaatgcatg
     1441 ttatttctgt gggtttttta attttcatct caactgggct tacaatttta acataataaa
     1501 aatcaaatca aatcaaatca ggcccttatt tgtcacatac actcttgtat actgaaataa
     1561 actaaatgtt gagaaaaatt taaatacacc aaaataacgc ataccttttt tgtaattacc
     1621 taagtaaaac atttagttcc ttttttaagt gactcaagat tgtaatctat taattgtaaa
     1681 agataaaact agtgatcaat ctgacgtgct tctatgcagc ctgtgagatt aattgtgatg
     1741 tttaattatt aaacaattta aatggatcaa acatttcaca tgtgcttatt gtgtccaaca
     1801 aaaccagcag aagcacttaa aaatattgca tgacaacatg agtttttaat gtaaattata
     1861 tatacttcct aaacgaagtt aatcatattt tctaaaataa taatgtcatt gaaatattta
     1921 aataaatgat aanctttagt tgattcaaca taaaattgaa agcaataagg actttttttt
     1981 ttttacagcg gtgaagcaaa agcntgnaaa taatcatata aatataataa ataaataaat
     2041 aaaataatgg ttcttggtgc aaatatgaac actttaaata aatatagaat aaatatgatc
     2101 aataagtcat aacgacgtgt tttatgttga ttataattta aacatgtttc taacgcaagt
     2161 tttaaaaact gttttaaagt attattttaa gttataagat caatttgact caacattttg
     2221 tagtgaaaat atgttgtgta aaattaatat aatatacact ataaaaatgc aaggttccac
     2281 acaattcatt catgttgtcc caacacaaat cgattaagtt aacaaatata tgtggattga
     2341 acataaaaaa ttaagttgtc ccaatgaaat ctcaagaatt gtgttgtttc agctcatttt
     2401 aaataagtag ttttaacaag taaaaaataa catatttttg agtgtgtata tatatatatt
     2461 tatatattac aaaatataat aatataaaat gtaaaagata taaaccatgc tacatatatt
     2521 taagagctat tttgcacatt acttttttgc aagccatttg agtgtttttt tacaaatata
     2581 tattagctaa atatacaata tcttaaatat acaattaatc agacatgcaa aaaatgcatt
     2641 taacctacat aaaaatacat gatcaacata tgttttatgc tgattacaac taaatcaaac
     2701 gtaatgaaat taatttataa aagcgattta aatatgtcaa tattttaagt taaaatcact
     2761 tgaaacctac tataaatgaa cttaaattta aaaaatctcc tttaattcca atgcagcact
     2821 gttgtttttt cagctgagga cccccggtgc agatggttgg cccccggccc gctattctaa
     2881 ggaagccttt cttaatttta gggcggcgct gcatgcgctc agatcagatt cacgggtgga
     2941 gagggttatt ttctcttttt tattttttct tgcctgctca tatttgggcc tgtgtgaacc
     3001 ccaggctccg cctcggcccc caggagccgc tcttatttaa aggggatcct cgccttcagt
     3061 cttcgtccca gtggaagcat cagtcttgtg ataacaactt ggtgagtcct acagaccagc
     3121 gctcgactct aggctttaaa tgcattgatt tctctggatt agctaccggg agagttttaa
     3181 ctttgcttat tagtcttgtg cttgaactgc taggtttaaa gagggttttc tctaatgcat
     3241 gcatgaagcg acagctgaaa accagaaata gagcgtttgt cctcctgtta aactctagcc
     3301 tttctttggc atttgtcagt ccgagtcttc agctgccggc aggagattgg gtttcagtaa
     3361 cacttgctga gcacaaacta cacgtgcttc cttcagattt aattcattat taatcattct
     3421 agatttaaat gatcttctta ctcattgctg attattttag ccttgcggag cagattaaat
     3481 gagcacgtgt ttattaagat tgatttaaat ataattaata tttcaccact gtagttttaa
     3541 aattgccttc taagtcatgg ttgattttta ataggtatcg gctctaatta gattactatt
     3601 ttggttttta aatgttattt ctatttcatt tgtaggctcg tttttaatgt gattttatta
     3661 gattctagtt ttaaaatcat cttttagcct tactgagcag actgaatgaa catgtttcat
     3721 taagattaac ttcattatag ttaattattt aatcattcta gtggattata aatcagctct
     3781 atttaaattg attgtcgtgt ttatttctat ttgatttttg attatttttt aaggtcgttt
     3841 ttatgtgatt tttccccgat tctaatttta aactaattgt ttgattttta tatgattcga
     3901 ctctttttaa atgaattgtt ttgctttttt tattgagcat atatatatat tatacaattt
     3961 caatcaacaa ttgttgtgct ttttaaatgt tgtttctatt tcatttctag ggtcgatttt
     4021 attcgattct agttttaaaa tcgtgatttc taactcctaa ccttactgag cagactgaat
     4081 gaacatgttt catcaagatt aacttcatta tagttaatta ttgaatcggg ctagtggttt
     4141 ataaatcagc tctattataa attaattgtc atcattattt ctatttaatt tttattattt
     4201 ttttaggatt atttttaagg gatttttttc acgttctagt tttaaactca atgctcatta
     4261 ctgctcaccc ttgctgagca gtctttcgtt aagattaaat taattaataa cccattctag
     4321 ttttaaaaca gtgtcttaag gcatagctga tattttacaa gtttactatt taatatatct
     4381 ccatttcttt ttttagggtc gatttcatct gatttgatta atttttatta tcttcactaa
     4441 gctttttgtt ctaggtttta ttctcagtat tattactgtt atatttgaac tgaatttctc
     4501 ctcatagttt attcatttta tttatttatt ttttgtttat tcgctatttt gtattttagt
     4561 ttttaattag aaattttatt agaatcgttt tgaactagtt tttatgcact ctaacattaa
     4621 tattaaatca taactattat ttttattata atttatttgt gtttatcttt ttgtgtagct
     4681 ttcatgtgtt tttattttag ttttattatg tttgcattga attttttgga ctagtttata
     4741 taacgatata cagaaaagcc atgacaacaa atgtttttat aatacctatt aagtataagc
     4801 taatcagatt tccactattc tgttcaaagt taacattatt agccattctg atggatcatt
     4861 tgacatttgt catatgtctt tatgaacact gctctttcag atgccacttt taaagctgtt
     4921 ttttacagta aatgaacgtt tattattatg acattatttc ctaaggcctc tagagtacag
     4981 caggatgtgg cgtgtgcatt actgtgtaat tatgctgact gtaaatgtgc atttatagac
     5041 tgtcagcatg tctgactgtg cgctgttgtg atttgtcctg cacagaccaa ggcaagtttg
     5101 tacaaaaaag caggctgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa
     5161 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
     5221 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc
     5281 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca
     5341 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag
     5401 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     5461 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt
     5521 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg
     5581 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg
     5641 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt
     5701 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga
     5761 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagaga
     5821 cccagctttc ttgtacaaag tggtggcaag tccagtagaa ttcgcaattg accccaatga
     5881 gccaacttat tgtctttgca atcaagttag ttatggcgag atgataggat gcgacaacga
     5941 gcagtgccca atagagtggt tccatttcag ttgcgtatca ttgacatata aaccaaaggg
     6001 caaatggtat tgcccaaagt gcaggggtga caatgagaag acgatggaca agagtacaga
     6061 gaagacaaaa aaagagcgac gtgctcgtga gttcgccagt ccggtggagt tcgcgattga
     6121 cccgaacgag cccacgtact gcttgtgtaa tcaggtatcc tacggggaaa tgatcggatg
     6181 cgacaatgaa cagtgtccca tcgagtggtt tcacttttcc tgcgtgtctt tgacatataa
     6241 acctaaagga aaatggtact gcccaaagtg tagaggagat aatgaaaaga ctatggataa
     6301 gtctaccgag aagactaaaa aggaacgccg agcgagggag ttcgcaagtc ccgttgaatt
     6361 tgcgattgac cctaatgagc cgacttactg cctttgcaac caggtttcat acggtgagat
     6421 gatcggctgt gacaacgaac aatgcccaat agaatggttc cacttttcat gtgtgagtct
     6481 tacatataaa cccaagggga aatggtattg ccccaagtgt cgaggcgaca atgaaaaaac
     6541 gatggacaag agcacagaaa aaaccaaaaa agagagacgt gcgcggtaac aactttatta
     6601 tacatagttg ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat
     6661 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     6721 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6781 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     6841 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc ctctagatca
     6901 gatctgcgaa gatacggcca cgggtgctct tgatcctgtg gctgattttg gactgtgctg
     6961 ctcgcagctg ctgatgaatc acatacttcc tccattttct tccactgatt gactgttata
     7021 atttccctaa tttccaggtc aaggtgctgt gcattgtggt aatagatgtg acatgacgtc
     7081 acttccaaag gaccaatgaa catgtctgac caatttcata taatgtgaaa acgattttca
     7141 taggcagaat aaataacatt taaattaaac tgggcatcag cgcaattcaa ttggtttggt
     7201 aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt tgactgtaaa
     7261 taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta
     7321 aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag
     7381 tacagtaatc aagtaaaatt actcaagtac tttacacctc tggttcttga ccccctacct
     7441 tcagcaagcc cagcagatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     7501 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     7561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     8401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     9601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9661 ataggggttc cgcgcacatt tccccgaaaa gtg
//
