LOCUS Exported 5837 bp ds-DNA circular SYN 27-OCT-2021 DEFINITION Mammalian expression of zebrafish Cavin4b-mCherry. Parton lab clone HYO. ACCESSION . VERSION . KEYWORDS Cavin4b-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5837) TITLE Harriet's Cavin4 paper JOURNAL Unpublished REFERENCE 2 (bases 1 to 5837) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 27, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5837 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 77..1054 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1089..1107 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1089..1106 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 1162..1186 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1187..1196 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2148..2172) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2174..2881 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(2316..2334) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2584..2603 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2778..2801 /label=DsRed1-C /note="DsRed1, forward primer" protein_bind complement(2883..2903) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter complement(2921..2938) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2943..3077 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2967..2986) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3021..3040 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3190..3210) /label=T3 /note="T3 promoter, forward primer" promoter complement(3190..3208) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3229..3245) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3229..3245) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3242..3264) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3253..3269 /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(3277..3307) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3322..3343 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3460..3477) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3631..4219) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3711..3730) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4390..5250) /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 5013..5032 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5251..5355) /gene="bla" /label=AmpR promoter rep_origin complement(5381..5836) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5518..5539) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5730..5749 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg 121 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg 181 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg 241 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg 301 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg 361 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca 421 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata 481 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata 541 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat 601 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt 661 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac 721 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta 781 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac 841 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg 901 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg 961 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg 1021 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag 1081 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca 1141 tcgattcgaa ttcaaggatc aacaagtttg tacaaaaaag caggctgcca ccatggctga 1201 taaactggga ctggcaggtg tggcggacga gcccagcgcc atgaacatct tctctctgct 1261 ggagagggtg tccgggatca tagacaatgt tcaggcctgc cagcagcgga tggaggagag 1321 gcagctggag ctggagaaca ccgtcaagac catccaagca gatgtagtca agctgacgaa 1381 ggagcacacg gcgaccagtg gcaccgtgga gaaacttctg gagaaaacac gcaaggtcag 1441 ctgccacatc aaggacgtcc gcgtgcgggt ggagaagcag aacgttcgag tcaagaaggt 1501 ggaggaaacc caagtcgagc tgctgaataa gaacaagttc cgtgtcgtca tctaccaggg 1561 agataatgag gtcccagctg tggctggtcc taaagcctct ccgaagggag caatgggtgg 1621 agacgctgct gcagacccag atgcaccaga ggctccgctc cctgactcgg atgaggaata 1681 catggttgtt gaagaagccg attcttccac tgctgccaaa ctaaagaaga ccggcctgaa 1741 gcgcatcgaa agcctgaagc agaccttctc acgcgagaac atgactaaga ccaaagaaaa 1801 cctgggcacc aaggtcaaca agctgggcga gcgcatagta acagccgaac ggcgtgagaa 1861 aatccgccag tcaggagaac gactgaaaca gtctggcgag cgcttcaagg aaaccatcac 1921 aaaaaccgta ccagcaaagc tgaatctgaa gaaggaaagg acggtggctg agggtcagga 1981 gggggcagaa gggaccacag aaggaacagc acccgtccca ccaccgaagg gtcgtaaacc 2041 cagtcctgag gtggcctaca cagagccaac gcagaaacaa gaaggtccag atgagggcaa 2101 gggtgaggaa tcagaagtac cgatgtatga tatgaagcag ctttctgacc cagctttctt 2161 gtacaaagtg gctgtgagca agggcgagga ggacaacatg gccatcatca aggagttcat 2221 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga 2281 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg 2341 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta 2401 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa 2461 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc 2521 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga 2581 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc 2641 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca 2701 ctacgacgcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc 2761 ctacaacgtc aacatcaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga 2821 acagtacgag cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta 2881 accaacttta ttatacatag ttgcactcga gcctctagaa ctatagtgag tcgtattacg 2941 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt 3001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa 3061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg 3121 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag 3181 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt 3241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa 3301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 3361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 3421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 3481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 3541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 3601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 3661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 3721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 3781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 3841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 3901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 3961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 4081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 4321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 4381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 4441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 4501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 4561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 4621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 4681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 4741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 4801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 4861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 4921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 4981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 5041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 5101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 5161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 5221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 5281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 5341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt 5401 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 5461 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca 5521 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 5581 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg 5641 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 5701 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 5761 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg 5821 ccgctacagg gcgcgtc //