別稱 | pACYC-184 |
復(fù)制子 | p15A |
質(zhì)粒分類 | 大腸系列質(zhì)粒;大腸克隆質(zhì)粒;基因克隆質(zhì)粒 |
質(zhì)粒大小 | 4245bp |
原核抗性 | Chl,Tet |
克隆菌株 | DH5a |
培養(yǎng)條件 | 37度 |
5'測序引物 | Tcr-F(GAAGTCATGCGCCGGTTAAG) |
3'測序引物 | pBRrevBam-R (GGTGATGTCGGCGATATAGG) |
備注 | 豐富的酶切位點(diǎn) |
載體宿主 | 大腸桿菌 |
載體用途 | PCR模板 |
基因種屬 | 空載體 |
基因類型 | cDNA |
原核抗性 | Chl,Tet |
質(zhì)粒簡介 | pACYC184質(zhì)粒是一種原核表達(dá)載體,含有p15A復(fù)制起點(diǎn)的大腸桿菌質(zhì)粒克隆載體。這允許pACYC184在與ColE1相容性組的質(zhì)粒(例如pBR322,pUC19)的細(xì)胞中共存。它是一個(gè)低拷貝數(shù)載體,每個(gè)細(xì)胞約15個(gè)拷貝,但可以用壯觀霉素放大。由于cat基因的存在,氯霉素不能用于提高質(zhì)粒產(chǎn)量(阻礙菌體復(fù)制,不阻礙質(zhì)粒擴(kuò)增)。 pACYC184 is an E. coli plasmid cloning vector containing the p15A origin of replication (1-4). This allows pACYC184 to coexist in cells with plasmids of the ColE1 compatibility group (e.g., pBR322, pUC19). It is a low copy number vector, at about 15 copies per cell (5), but can be amplifed with spectinomycin. Chloramphenicol cannot be used for amplifcation due to the presence of the cat gene. |
質(zhì)粒序列 |
LOCUS Exported 4245 bp ds-DNA circular SYN 26-8-2015 DEFINITION synthetic circular DNA KEYWORDS Untitled 2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4245) TITLE Direct Submission JOURNAL Exported 2015-8-26 FEATURES Location/Qualifiers source 1..4245 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(220..322) /note="cat promoter" /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" rep_origin complement(848..1393) /direction=LEFT /note="p15A ori" /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin." promoter 1505..1533 /note="tet promoter" /note="E. coli promoter for tetracycline efflux protein gene" CDS 1581..2771 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /note="TcR" /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" CDS complement(3805..219) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" ORIGIN 1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt 61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt 121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga 181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga 241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt 301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc 361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat 421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt 481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg 541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact 601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa 661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc 721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc 781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa 841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc 901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc 961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc 1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac 1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt 1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt 1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg 1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt 1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc 1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca 1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc 1501 atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct 1561 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg 1621 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg 1681 atatcgtcca ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt 1741 tgatgcaatt tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc 1801 cagtcctgct cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac 1861 ccgtcctgtg gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg 1921 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg 1981 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt 2041 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc 2101 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccgatgccct 2161 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg 2221 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg 2281 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg 2341 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt 2401 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc 2461 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg 2521 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg 2581 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga 2641 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag 2701 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca 2761 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg 2821 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc 2881 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg 2941 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg 3001 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg 3061 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag 3121 tctggaaacg cggaagtccc ctacgtgctg ctgaagttgc ccgcaacaga gagtggaacc 3181 aaccggtgat accacgatac tatgactgag agtcaacgcc atgagcggcc tcatttctta 3241 ttctgagtta caacagtccg caccgctgtc cggtagctcc ttccggtggg cgcggggcat 3301 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc 3361 ggcagcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc 3421 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac 3481 acctacatct gtattaacga agcgctaacc gtttttatca ggctctggga ggcagaataa 3541 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca 3601 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag 3661 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg 3721 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg 3781 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg 3841 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat 3901 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg 3961 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg 4021 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc 4081 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta 4141 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg 4201 aacactatcc catatcacca gctcaccgtc tttcattgcc atacg |