• 规格：20μl质粒
• 启动子：CMV promoter
• 复制子：Ori，F1 ori
• 终止子：bGH poly(A) signal
• 质粒分类：基因文库质粒；大鼠基因质粒；大鼠真核质粒
• 质粒大小：10850bp
• 原核抗性：氨苄青霉素Amp
• 筛选标记：杀稻瘟素Blast
• 克隆菌株：大肠杆菌Top10F
• 表达宿主：293T等哺乳细胞
• 质粒简介：               CaV1.3e[8a,11,31b,Δ32,42a] mut质粒的参考文献：Neuronal Ca(V)1.3alpha(1) L-type channels activate at relatively hyperpolarized membrane potentials and are incompletely inhibited by dihydropyridines. 
                Contains exons 8a, 11, 31b and 42a.Deletion of exon 32. nt 3310 (GTG->GCG) [Val->Ala]; nt 731 (TCA->GGA) [Ser->Gly].
• 质粒图谱1： png/1-1FQG00214K9.png

质粒序列： 点击查看序列

LOCUS       Exported               10850 bp ds-DNA    circular SYN 17-八月-2017

KEYWORDS    CaV1.3e[8a,11,31b,Delta-32,42a] mut

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 10850)

FEATURES             Location/Qualifiers

     source          1..10850

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        235..614

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        615..818

                     /note="CMV promoter"

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     promoter        863..881

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    901..5832

                     /note="CaV1.3e[8a,11,31b,Delta-32,42a] mut-r"

                     /note="Rattus norvegicus voltage-gated calcium channel pore

                     forming subunit CaV1.3alpha1 mRNA, complete cds, 

                     alternatively spliced

                     GenBank: AF370009.1"

     misc_feature    5833..6679

                     /note="3'UTR"

     primer_bind     complement(6663..6679)

                     /note="SK primer"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     polyA_signal    6812..7036

                     /note="bGH poly(A) signal"

                     /note="bovine growth hormone polyadenylation signal"

     rep_origin      7082..7510

                     /direction=RIGHT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        7524..7853

                     /note="SV40 promoter"

                     /note="SV40 enhancer and early promoter"

     rep_origin      7704..7839

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     promoter        7901..7948

                     /note="EM7 promoter"

                     /note="synthetic bacterial promoter "

     CDS             7967..8365

                     /codon_start=1

                     /gene="Aspergillus terreus BSD"

                     /product="blasticidin S deaminase"

                     /note="Blast"

                     /note="confers resistance to blasticidin"

                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG

                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI

                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"

     polyA_signal    8523..8644

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     protein_bind    8717..8733

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /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(8741..8771)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    8786..8807

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(9095..9683)

                     /direction=LEFT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(9854..10714)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="AmpR"

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(10715..10819)

                     /gene="bla"

                     /note="AmpR promoter"

ORIGIN

        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg

       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg

      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc

      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt

      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata

      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc

      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc

      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt

      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt

      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca

      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg

      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc

      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg

      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca

      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc

      901 atgcagcatc aacggcagca gcaagaggac cacgcgaacg aggcaaacta tgcaagaggc

      961 accagactcc ctatttctgg tgaaggacca acttctcagc cgaacagctc caagcaaacc

     1021 gtcctgtctt ggcaagctgc aattgatgct gctagacagg ccaaggccgc ccagaccatg

     1081 agcacgtctg cacccccacc tgtaggatct ctctcccaaa gaaaacgtca gcaatacgcc

     1141 aagagcaaaa aacagggtaa ctcgtctaac agccgacctg cccgtgccct cttctgttta

     1201 tccctcaata accccatccg aagagcctgc attagtatag tggattggaa accatttgac

     1261 atatttatat tattggctat ttttgccaat tgtgtggcct tagctattta catcccgttc

     1321 cctgaagatg attctaattc aacaaatcat aacttggaaa aagtagaata tgccttcctg

     1381 attatcttta cagtggagac atttctgaag attatagcgt acggactgct gctgcatcct

     1441 aatgcttacg ttaggaacgg atggaactta ctggattttg ttatagtaat agtaggattg

     1501 tttagtgtaa ttttggaaca attaaccaaa gaaacagaag gcgggaacca ctctagtggc

     1561 aaatcaggag gctttgatgt caaagccctc cgtgccttcc gagtgttaag accgcttcga

     1621 cttgtatcat cagtgcccag tttacaagtt gttctgaact ccattataaa agccatggtc

     1681 cccctcctcc acatagccct tttggtgtta ttcgtaatta tcatctatgc gattataggg

     1741 ttggaacttt ttattggaaa aatgcacaaa acctgttttt ttgctgactc agatatagta

     1801 gctgaagagg acccagctcc atgcgcgttc tcagggaatg gacgccagtg tgccgccaat

     1861 ggcacggaat gtaggagcgg ctgggtcggc cccaatggag gcatcactaa ttttgacaac

     1921 tttgcctttg ccatgctcac tgtgttccag tgcatcacca tggagggctg gacagatgtg

     1981 ctctactggg ttaatgatgc gataggatgg gagtggccat gggtgtattt tgttagtctg

     2041 atcatccttg gctcattttt cgtccttaac ctggttcttg gtgtccttag tggagaattc

     2101 tcaaaggaaa gagagaaggc taaagcacgt ggagatttcc agaagcttcg ggagaagcag

     2161 cagctggaag aggatctcaa gggctacttg gactggatca cccaggcaga agacatagat

     2221 cctgagaatg aagaggaggg tggagaggag ggcaaacgaa acactagcat gcccaccagt

     2281 gagaccgaat ctgtgaacac agagaacgta agtggtgaag gcgagaccca gggatgctgt

     2341 ggaagtctct ggtgctggtg gaaaagaaga ggcgcggcca agactgggcc ctctgggtgt

     2401 cggcggtggg gccaagccat ctcgaaatcc aagctcagcc gtcggtggcg tcgctggaac

     2461 cggttcaatc gcaggagatg cagggcagct gtgaagtctg tcacgtttta ctggctggtg

     2521 atcgttctgg tgtttctcaa cactctgacg atttcctctg agcactacaa tcagccagac

     2581 tggctgacgc agattcaaga tatagcgaac aaagtgctct tggctttgtt cacctgtgag

     2641 atgctggtga agatgtacag cctgggcctc caggcctact tcgtctctct cttcaaccgc

     2701 ttcgattgct ttgtggtgtg tgggggaatc accgaaacca tcctggtgga actggagctc

     2761 atgtctcccc tgggcgtctc tgtgttccgg tgtgtacgtc tcctgaggat cttcaaagtg

     2821 accaggcact ggacttccct gagcaacttg gtggcgtccc tgttaaactc catgaagtcc

     2881 atcgcctctc tgctgttgct gctcttcctc ttcatcatca tcttctccct gctggggatg

     2941 cagctgttcg gagggaagtt caatttcgat gagacacaga ccaagcgaag caccttcgac

     3001 aacttcccac aggcgctcct gactgtgttc cagatcctga cgggcgaaga ctggaatgcg

     3061 gtgatgtatg acggcatcat ggcctacgga ggcccgtcct cctctggaat gattgtctgt

     3121 atttacttca tcatcctttt catctgtggt aactatatcc tgctgaacgt cttcttggcc

     3181 attgctgtag acaatttggc tgatgctgaa agtctgaaca ctgctcagaa agaggaagcc

     3241 gaagaaaaag aaaggaaaaa aattgccaga aaagaaagcc tagaaaacaa aaaaaacaac

     3301 aaaccagagg tcaaccagat agccaacagt gacaacaagg ttactattga tgactatcaa

     3361 gaggaggctg aagataagga cc