LOCUS Exported 5657 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Baculovirus expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS KCNK2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5657) TITLE SGC Oxford REFERENCE 2 (bases 1 to 5657) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5657 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 2..457 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(89..108) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 299..320 /label=F1ori-F /note="F1 origin, forward primer" promoter 484..588 /gene="bla" /label=AmpR promoter CDS 589..1449 /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(807..826) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1620..2208 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2109..2128 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2362..2379 /label=L4440 /note="L4440 vector, forward primer" mobile_element 2511..2735 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(2802..3335) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(3524..3552) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" promoter 3904..3995 /gene="polh from Autographa californica" /label=polyhedrin promoter /note="promoter for the baculovirus polyhedrin gene" primer_bind 3920..3937 /label=Polyhedrin-F /note="Polyhedrin promoter, forward primer" CDS 4915..4935 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 4936..4965 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 4966..4989 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 5147..5281 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5184..5203) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5238..5257 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" mobile_element 5310..5475 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" ORIGIN 1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat 481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 1021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 1081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 1141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 1201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 1261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 1321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 1381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 1441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc 1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 1981 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg 2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 2221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 2281 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 2341 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg 2401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct 2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga 2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag 2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt 2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga 2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac 2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg 2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg 2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca 2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact 3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc 3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta 3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct 3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg 3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg 3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca 3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa 3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa 3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca 3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac 3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc 3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg 3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt 3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt 3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa 3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt 3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca 4021 ccatcgggcg cggatctcct taagaaggag atatactatg caggtttaag aaggagatat 4081 actatgccca cagtgcttgc ttcccgggtg gagagtgaca cgaccattaa tgttatgaaa 4141 tggaagacgg tctccacgat attcctggtg gttgtcctct atctgatcat cggagccacc 4201 gtgttcaaag cattggagca gcctcatgag atttcacaga ggaccaccat tgtgatccag 4261 aagcaaacat tcatatccca acattcctgt gtcaattcga cggagctgga tgaactcatt 4321 cagcaaatag tggcagcaat aaatgcaggg attataccgt taggaaacac ctccaatcaa 4381 atcagtcact gggatttggg aagttccttc ttctttgctg gcactgttat tacaaccata 4441 ggatttggaa acatctcacc acgcacagaa ggcggcaaaa tattctgtat catctatgcc 4501 ttactgggaa ttcccctctt tggttttctc ttggctggag ttggagatca gctaggcacc 4561 atatttggaa aaggaattgc caaagtggaa gatacgttta ttaagtggaa tgttagtcag 4621 accaagattc gcatcatctc aacaatcata tttatactat ttggctgtgt actctttgtg 4681 gctctgcctg cgatcatatt caaacacata gaaggctgga gtgccctgga cgccatttat 4741 tttgtggtta tcactctaac aactattgga tttggtgact acgttgcagg tggatccgat 4801 attgaatatc tggacttcta taagcctgtc gtgtggttct ggatccttgt agggcttgct 4861 tactttgctg ctgtcctgag catgattgga gattggctcc gagtgatatc tgcagagaac 4921 ctctacttcc aatcgcacca tcatcaccat caccatcacc accatgatta caaggatgac 4981 gacgataagt gaggatccga attcgagctc cgtcgacaag cttgtcgaga agtactagag 5041 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 5101 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 5161 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 5221 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat 5281 ctgatcactg cttgagccta ggagatccga accagataag tgaaatctag ttccaaacta 5341 ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc catctatttt 5401 gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta 5461 ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc 5521 caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt 5581 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca 5641 gcctgaatgg cgaatgg //