LOCUS Exported 6764 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Baculovirus expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS ANO10 (TMEM16KA-c001) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6764) TITLE SGC Oxford April 2018 plasmids REFERENCE 2 (bases 1 to 6764) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6764 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 126..217 /gene="polh from Autographa californica" /label=polyhedrin promoter /note="promoter for the baculovirus polyhedrin gene" primer_bind 142..159 /label=Polyhedrin-F /note="Polyhedrin promoter, forward primer" CDS 2263..2283 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 2284..2313 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 2314..2337 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 2495..2629 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2532..2551) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2586..2605 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" mobile_element 2658..2823 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" rep_origin 3007..3461 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3094..3113) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3303..3324 /label=F1ori-F /note="F1 origin, forward primer" promoter 3487..3591 /gene="bla" /label=AmpR promoter CDS 3592..4452 /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 complement(3810..3829) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4623..5211 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5112..5131 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5365..5382 /label=L4440 /note="L4440 vector, forward primer" mobile_element 5516..5740 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" promoter complement(6510..6538) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" ORIGIN 1 gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt 61 ttgttcgccc aggactctag ctatagttct agtggttggc tacgtatact ccggaatatt 121 aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg 181 ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc 241 caccatcggg cgcggatcta cttaagaagg agatatacta tgaaagtgac gttatcagct 301 ttggatactt ctgagagttc tttcacacct ttggtggtca tagaacttgc tcaggatgtc 361 aaagaagaaa ccaaagaatg gctgaaaaac agaattatag ctaaaaaaaa agatggaggt 421 gcccagttgt tgtttagacc attgttaaat aaatatgaac aagaaacact agaaaatcag 481 aacttatatc ttgttggtgc ctccaagatt agaatgttac taggggcaga agcagtggga 541 ttggtaaaag agtgcaatga taacaccatg agagccttca catacagaac cagacagaac 601 ttcaaaggtt ttgatgataa caatgatgat ttcctgacaa tggcagaatg tcaattcatt 661 atcaaacatg aacttgaaaa tcttagagct aaagatgaaa aaatgatccc tggttaccct 721 caggcaaagt tgtatccagg aaaatcattg ttgagaagat tgctcacgtc tggcatcgtg 781 attcaggtgt ttccactgca tgacagtgaa gccctgaaga agcttgagga cacctggtac 841 actcggtttg ctttgaagta tcagcccata gacagtattc gtggctactt tggggaaaca 901 attgctctgt actttggatt tttggagtat ttcacttttg cattaatccc catggctgtc 961 attgggttac cttactactt gtttgtgtgg gaagactatg acaagtacgt gatctttgcc 1021 tcgttcaacc tcatctggtc cacggtgatt ctggaactgt ggaagcgtgg ctgtgccaac 1081 atgacctaca ggtgggggac actgctcatg aagagaaagt ttgaggagcc ccggccagga 1141 tttcatggtg tcttgggtat caattccatc actgggaagg aggagcctct gtaccccagc 1201 tacaagagac agttgcgcat ttacctggtc tccctgccat tcgtgtgcct ctgcctctat 1261 ttctcactgt atgtcatgat gatttacttc gacatggagg tttgggcctt gggtctacat 1321 gagaacagcg ggtctgagtg gaccagtgtc ctgttgtatg tgcccagcat catctatgcc 1381 attgtgattg agatcatgaa tcgtctctat cgatatgctg ccgagttttt aacttcatgg 1441 gagaatcaca gattggaatc tgcctatcag aaccatctaa ttctgaaagt tttagtgttc 1501 aacttcctca attgctttgc ctcactcttc tatattgcct ttgtcttgaa agatatgaag 1561 cttttgcgcc agagcttggc cactctccta attacctccc agatcctcaa ccaaattatg 1621 gaatcttttc ttccttattg gctccaaagg aagcatggtg tgcgggtgaa gaggaaggtg 1681 caggctttaa aggcagacat tgatgctaca ttatatgaac aagtcatcct ggaaaaagaa 1741 atgggaactt atttgggcac ctttgatgat tacttggagt tattcctgca gtttggttat 1801 gtgagccttt tctcctgtgt ttacccatta gcagctgcct ttgctgtgtt aaataacttc 1861 actgaagtaa attcagatgc cttaaaaatg tgcagggtct tcaaacgtcc attctcagaa 1921 ccttcagcca atattggtgt gtggcagttg gcttttgaaa cgatgagtgt tatatctgtg 1981 gtcactaact gtgcgctgat tggaatgtca ccacaagtga atgcagtctt tccagaatca 2041 aaagcagacc tcattttgat tgtagtagca gtggagcacg cactcctggc tttaaagttt 2101 atacttgcat ttgccatacc tgataagcca cggcatatcc agatgaaact agccagactg 2161 gaatttgagt ctttggaggc actcaagcag cagcaaatga agctcgtgac cgagaacctg 2221 aaggaggaac caatggaaag cgggaaggag aaggcaaccg cagagaacct ctacttccaa 2281 tcgcaccatc atcaccatca ccatcaccac catgattaca aggatgacga cgataagtga 2341 ggatccgaat tcgagctccg tcgacaagct tgtcgagaag tactagagga tcataatcag 2401 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 2461 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 2521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 2581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct 2641 tgagcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt 2701 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc 2761 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc 2821 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg 2881 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct 2941 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg 3001 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 3061 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa 3301 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3361 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3421 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga 3481 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 3541 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 3601 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 3661 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 3721 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 3781 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 3841 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 3901 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 3961 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 4021 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 4081 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 4141 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 4201 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 4261 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 4321 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 4381 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 4441 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 4501 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 4561 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 4621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 4681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 4741 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 4801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 4861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 4921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 4981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5101 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5161 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 5221 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 5281 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 5341 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 5401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt 5461 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg 5521 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa 5581 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg 5641 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt 5701 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac 5761 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac 5821 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca 5881 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg 5941 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg 6001 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt 6061 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct 6121 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta 6181 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag 6241 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc 6301 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt 6361 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag 6421 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt 6481 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact 6541 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg 6601 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc 6661 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct 6721 ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccg //