LOCUS Exported 9906 bp ds-DNA circular SYN 13-SEP-2021 DEFINITION Zebrafish muscle specific expression of Cavin4b-Clover. Parton lab clone DQZ. ACCESSION . VERSION . KEYWORDS acta1-Cavin4b-Clover SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9906) AUTHORS Xiong Z, Lo HP, McMahon KA, Martel N, Jones A, Hill MM, Parton RG, Hall TE TITLE In vivo proteomic mapping through GFP-directed proximity-dependent biotin labelling in zebrafish. JOURNAL Elife. 2021 Feb 16;10. pii: 64631. doi: 10.7554/eLife.64631. PUBMED 33591275 REFERENCE 2 (bases 1 to 9906) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9906 /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 5074..5098 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 5099..5108 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(6060..6084) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 6086..6802 /codon_start=1 /product="monomeric derivative of the bright green-yellow fluorescent protein Clover" /label=mClover2 /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(6128..6149) /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(6803..6823) /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(6831..6847) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" polyA_signal 6863..6997 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6887..6906) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6941..6960 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(7669..7685) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7669..7685) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(7722..7742) /label=T3 /note="T3 promoter, forward primer" promoter complement(7722..7740) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7761..7777) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7774..7796) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7785..7801 /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(7809..7839) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7854..7875 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7992..8009) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8163..8751) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8243..8262) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8922..9782) /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 9545..9564 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9783..9887) /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 atgctcttgg 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 ttttaagtac ctaaatgtgt tcaaaagtaa 1321 agtatgttat tgtttttttt ttttaatgtt agctaagctt taaattgtaa tatattactt 1381 ttaaaagtga ctttttaaaa atgtaaataa aaaagtaact ataaatgtgt aatgcatgtt 1441 atttctgtgg gtttttaatt ttcatctcaa ctgggcttac aattttaaca taataaaaat 1501 caaatcaaat caaatcaggc ccttatttgt cacatacact cttgtatact gaaataaact 1561 aaatgttgag aaaaatttaa atacaccaaa ataacgcata ccttttttgt aattacctaa 1621 gtaaaacatt tagttccttt tttaagtgac tcaagattgt aatctattaa ttgtaaaaga 1681 taaaactagt gatcaatctg acgtgcttct atgcagcctg tgagattaat tgtgatgttt 1741 aattattaaa caatttaaat ggatcaaaca tttcacatgt gcttattgtg tccaacaaaa 1801 ccagcagaag cacttaaaaa tattgcatga caacatgagt ttttaatgta aattatatat 1861 acttcctaaa cgaagttaat catatttcta aaataataat gtcattgaaa tatttaaata 1921 aatgataagc tttagttgat tcaacataaa attgaaagca ataaggactt ttttttttta 1981 cagcggtgaa gcaaaagcat gtaataatca tataatataa taaataaata aataaataat 2041 ggctcttggt gcaaatatga acactttaaa taaatataga ataaatatga tcaataagtc 2101 ataacgatgt gttttatgtt gattataatt taaacatgtt tctaacgcaa gttttaaaaa 2161 ctgttttaaa gtattatttt aagttataag atcaatttga ctcaacattt tgtagtgaaa 2221 atatgttgtg taaaattaat ataatataca ctataaaaat gcagggttcc acacaataca 2281 ttaatgttgt cccaacacaa atcgattaag ttaacaaata tatgtggatt gaacataaaa 2341 aattaagttg tcccaatgaa atctcaagaa ttgtgttgtt tcagctcatt ttaaataagt 2401 agttttaaca agtaaaaaat aacatatttt tgagtgtgta tatatatata ttacaaaata 2461 taataatata aaatgtaaaa gatataaacc atgctacata tatttaagag ctattttgca 2521 cattactttt ttgcaagcca ttttagtgtt ttttttacaa atatatatta gctaaatata 2581 caatatctta aatatacaat taatcagaca tgcaattaat gcatttaacc tacataaaaa 2641 tacatgatca acatatgttt tatgctgatt acaactaaat caaacgtaat gaaattaatt 2701 tataaaagcg atttaaatat gtcaatattt taagttaaaa tcacttgaaa cctactataa 2761 atgaacttaa attaaaaaaa tctcctttaa ttccaatgca acagtgttgt tttttcagct 2821 gaggaccccc ggtgcagatg gttggccccc ggcccgctat tctaaggaag cctttcttaa 2881 ttttagggcg gcgctgcatg cgctcagatc agattcacgg gtggagaggg ttattttctc 2941 ttttttattt tttcttgcct gctcatattt gggcctgtgt gaaccccagg ctccgcctcg 3001 gcccccagga gccgctctta tttaaagggg atcctcgcct tcagtcttcg tcccagtgga 3061 agcatcagtc ttgtgataac aacttggtga gtcctacaga ccagcgctcg actctaggct 3121 ttaaatgcat tgatttctct ggattagcta ccgggagagt tttaactttg cttattagtc 3181 ttgtgctcga actgctaggt ttaaagagtg ttttctctaa tgcatgcatg aagcgacagc 3241 tgaaaaccag aaatagagcg tttgtcctcc tgttaaactc tagcctttct ttggcatttg 3301 tcagtccgag tcttcagctg ccggcaggag attgggtttc agtaacactt gctgagcaca 3361 aactacacgt gctttcttca gatttaattc attattaatc attctagatt taaatgatgt 3421 tcttactcat tgctgattat tttagccttg cggagcagat taaatgagca cgtgtttcat 3481 taagattgat ttaaatataa ttaatatttc accactgtag ttttaaaatt gccttctaag 3541 tcatggttga tttttaatag gtatcggctc taattagatt actattttgg tttttaaatg 3601 ttatttctat ttcatttgta ggctcgtttt taatgtgatt ttattagatt ctagttttaa 3661 aatcatctct tagccttaat gagcagactg aatgaacatg tttcattaag attaacttca 3721 ttataatgaa ttatttaatc attctagtgg gttataaatc agctctattt aaattaattg 3781 tcgtgtttat ttctatttaa tttttgatta ttttttcagg gtcgttttta tgtgattttt 3841 tcccgtttct aattttaaac taattgtttg atttttatat gattcgactc tttttaaatg 3901 aattgttttg ctttttttat tgagcatata tatattatac aatttcaatc aacaattgtt 3961 gtgcttttta aatgttgttt ctatttcatt tctagggtcg attttattcg attctagttt 4021 taaaatcgtg atttttaact tctaacctta ctgagcagac tgaatgaaca tgtttcatca 4081 agattaactt cattatagtt aattattgaa tcgtgctagt ggtttataaa tcagctctat 4141 ttaaattaat catcatcatt atttctattt aatttttatt atttctttag gattattttt 4201 atgtgatttt tttcacgttc tagttttaaa ctcaatgctc attactgctc acccttgctg 4261 agcagtcttt cgttaagatt aaattaatta ataaccattc tagttttaaa acagtgtctt 4321 aaggcatagc tgatatttta caagtttact atttaatata tctcctttct ttttttaggg 4381 tcgatttcat ctgatttgat taatttttat tatcttcact aagctttttg ttctaggttt 4441 tattctcagt attattactg ttatatttga actgaatttc tcctcatagt ttattcattt 4501 tatttattta ttttttgttt attcgctatt ttgtatttta gtttttaatt agaaatttta 4561 ttagaatcgt tttgaactag tttttatgca ctctaacatt aatattaaat cataactatt 4621 atttttatta taatttattt gtgtttatct ttttgtgtag ctttcatgtg tttttatttt 4681 agttttatta tgtttgcatt gaattttttg gactagttta tataacgata tacagaaaag 4741 ccatgacaac aaatgttttt ataataccta ttaagtataa gctaatcaga tttccactat 4801 tctgttcaaa gttaacatta ttagccattc tgatggatca tttgacattt gtcatatgtc 4861 tttatgaaca ctgctctttc agatgccact tttaaagctg ttttttacag taaatgaacg 4921 tttattatta tgacattatt tcctaaggcc tctagagtac agcaggatgt ggcgtgtgca 4981 ttaccgtgta attatgctga ctgtaaatgt gcatttatag actgtcagca tgtctgactg 5041 tgcgctgttg tgattggtcc tgcacagacc aaggcaagtt tgtacaaaaa agcaggctgc 5101 caccatggct gataaactgg gactggcagg tgtggcggac gagcccagcg ccatgaacat 5161 cttctctctg ctggagaggg tgtccgggat catagacaat gttcaggcct gccagcagcg 5221 gatggaggag aggcagctgg agctggagaa caccgtcaag accatccaag cagatgtagt 5281 caagctgacg aaggagcaca cggcgaccag tggcaccgtg gagaaacttc tggagaaaac 5341 acgcaaggtc agctgccaca tcaaggacgt ccgcgtgcgg gtggagaagc agaacgttcg 5401 agtcaagaag gtggaggaaa cccaagtcga gctgctgaat aagaacaagt tccgtgtcgt 5461 catctaccag ggagataatg aggtcccagc tgtggctggt cctaaagcct ctccgaaggg 5521 agcaatgggt ggagacgctg ctgcagaccc agatgcacca gaggctccgc tccctgactc 5581 ggatgaggaa tacatggttg ttgaagaagc cgattcttcc actgctgcca aactaaagaa 5641 gaccggcctg aagcgcatcg aaagcctgaa gcagaccttc tcacgcgaga acatgactaa 5701 gaccaaagaa aacctgggca ccaaggtcaa caagctgggc gagcgcatag taacagccga 5761 acggcgtgag aaaatccgcc agtcaggaga acgactgaaa cagtctggcg agcgcttcaa 5821 ggaaaccatc acaaaaaccg taccagcaaa gctgaatctg aagaaggaaa ggacggtggc 5881 tgagggtcag gagggggcag aagggaccac agaaggaaca gcacccgtcc caccaccgaa 5941 gggtcgtaaa cccagtcctg aggtggccta cacagagcca acgcagaaac aagaaggtcc 6001 agatgagggc aagggtgagg aatcagaagt accgatgtat gatatgaagc agctttctga 6061 cccagctttc ttgtacaaag tggtggtgag caagggcgag gagctgttca ccggggtggt 6121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga 6181 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa 6241 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag 6301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 6361 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt 6421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 6481 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat 6541 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc acaacgttga 6601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 6661 cgtgctgctg cccgacaacc actacctgag ccatcagtcc gccctgagca aagaccccaa 6721 cgagaagcgc gatcacatgg tcctgctgga gttcgtaacc gccgccggga tcactctcgg 6781 catggacgag ctgtacaagt aacaacttta ttatacatag ttgataattc actggccgtc 6841 gttttacggt accatcgatg atgatccaga catgataaga tacattgatg agtttggaca 6901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc 6961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt 7021 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa 7081 atgtggtatg gctgattatg atcctctaga tcagatctgc gaagatacgg ccacgggtgc 7141 tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact 7201 tcctccattt tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc 7261 tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct 7321 gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta 7381 aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg 7441 atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt 7501 ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa 7561 gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag 7621 tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga tccactagtt 7681 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt 7741 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 7801 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 7861 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 7921 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 7981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 8041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 8101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 8161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 8221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 8281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 8341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 8401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 8461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 8521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 8581 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 8641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 8701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 8761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta aggcattttg 8821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 8881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 8941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 9001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 9061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 9121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 9181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 9241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 9301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 9361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 9421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 9481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 9541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 9601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 9661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 9721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 9781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 9841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 9901 aaagtg //