| 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；其它基因质粒；鱼源基因质粒 |
|---|
| 质粒大小: | 8058bp |
|---|
| 质粒标签: | N-3xFLAG，N-SV40 NLS，C-FokI cleavage domain |
|---|
| 原核抗性: | 氨苄青霉素Amp（100μg/ml） |
|---|
| 筛选标记: | 杀稻瘟菌素blast |
|---|
| 克隆菌株: | XL1 Blue等大肠杆菌 |
|---|
| 培养条件: | 37℃，有氧，LB |
|---|
| 表达宿主: | 293T等哺乳细胞 |
|---|
| 备注: | 高拷贝质粒 |
|---|
TAL3393质粒是一个表达斑马鱼基因的真核表达质粒，CMV promoter启动ZebrafishCommunity-vasa-right基因的表达。
LOCUS Exported 8058 bp ds-DNA circular SYN 16-八月-2017
KEYWORDS TAL3393
FEATURES Location/Qualifiers
source 1..8058
/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"
CDS 907..972
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 979..999
/codon_start=1
/locus_tag="
"
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/note="
"
/protein_id="
"
/translation="PKKKRKV"
misc_feature 1020..3312
/note="ZebrafishCommunity-vasa-right"
CDS 3319..3906
/codon_start=1
/product="nonspecific DNA cleavage domain of the FokI
endonuclease (Li et al., 1992)"
/note="FokI cleavage domain"
/note="must dimerize to be catalytically active"
/protein_id="
"
/translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
polyA_signal 4019..4243
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 4289..4717
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4731..5061
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 4912..5047
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 5109..5156
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 5175..5573
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/note="Blast"
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 5731..5852
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
protein_bind 5925..5941
/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(5949..5979)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5994..6015
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6303..6891)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7062..7922)
/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(7923..8027)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
961 gacgatgaca agatggcccc caagaagaag aggaaggtgg gcattcaccg cggggtacct
1021 atggtggact tgaggacact cggttattcg caacagcaac aggagaaaat caagcctaag
1081 gtcaggagca ccgtcgcgca acaccacgag gcgcttgtgg ggcatggctt cactcatgcg
1141 catattgtcg cgctttcaca gcaccctgcg gcgcttggga cggtggctgt caaataccaa
1201 gatatgattg cggccctgcc cgaagccacg cacgaggcaa ttgtaggggt cggtaaacag
1261 tggtcgggag cgcgagcact tgaggcgctg ctgactgtgg cgggtgagct tagggggcct
1321 ccgctccagc tcgacaccgg gcagctgctg aagatcgcga agagaggggg agtaacagcg
1381 gtagaggcag tgcacgcctg gcgcaatgcg ctcaccgggg cccccttgaa cctgacccca
1441 gaccaggtag tcgcaatcgc gtcacatgac gggggaaagc aagccctgga aaccgtgcaa
1501 aggttgttgc cggtcctttg tcaagaccac ggccttacac cggagcaagt cgtggccatt
1561 gcatcccacg acggtggcaa acaggctctt gagacggttc agagacttct cccagttctc
1621 tgtcaagccc acgggctgac tcccgatcaa gttgtagcga ttgcgtcgaa cattggaggg
1681 aaacaagcat tggagactgt ccaacggctc cttcccgtgt tgtgtcaagc ccacggtttg
1741 acgcctgcac aagtggtcgc catcgcctcc aatattggcg gtaagcaggc gctggaaaca
1801 gtacagcgcc tgctgcctgt actgtgccag gatcatggac tgaccccaga ccaggtagtc
1861 gcaatcgcgt cgaacattgg gggaaagcaa gccctggaaa ccgtgcaaag gttgttgccg
1921 gtcctttgtc aagaccacgg ccttacaccg gagcaagtcg tggccattgc aagcaatggg
1981 ggtggcaaac aggctcttga gacggttcag agacttctcc cagttctctg tcaagcccac
2041 gggctgactc ccgatcaagt tgtagcgatt gcgtcgcatg acggagggaa acaagcattg
2101 gagactgtcc aacggctcct tcccgtgttg tgtcaagccc acggtttgac gcctgcacaa
2161 gtggtcgcca tcgccagcca tgatggcggt aagcaggcgc tggaaacagt acagcgcctg
2221 ctgcctgtac tgtgccagga tcatggactg accccagacc aggtagtcgc aatcgcgtca
2281 aacggagggg gaaagcaagc cctggaaacc gtgcaaaggt tgttgccggt cctttgtcaa
2341 gaccacggcc ttacaccgga gcaagtcgtg gccattgcaa ataataacgg tggcaaacag
2401 gctcttgaga cggttcagag acttctccca gttctctgtc aagcccacgg gctgactccc
2461 gatcaagttg tagcgattgc gtcgaacatt ggagggaaac aagcattgga gactgtccaa
2521 cggctccttc ccgtgttgtg tcaagcccac ggtttgacgc ctgcacaagt ggtcgccatc
2581 gccaacaaca acggcggtaa gcaggcgctg gaaacagtac agcgcctgct gcctgtactg
2641 tgccaggatc atggactgac cccagaccag gtagtcgcaa tcgcgtcaca tgacggggga
2701 aagcaagccc tggaaaccgt gcaaaggttg ttgccggtcc tttgtcaaga ccacggcctt
2761 acaccggagc aagtcgtggc cattgcaagc aatgggggtg gcaaacaggc tcttgagacg
2821 gttcagagac ttctcccagt tctctgtcaa gcccacgggc tgactcccga tcaagttgta
2881 gcgattgcga ataacaatgg agggaaacaa gcattggaga ctgtccaacg gctccttccc
2941 gtgttgtgtc aagcccacgg tttgacgcct gcacaagtgg tcgccatcgc cagccatgat
3001 ggcggtaagc aggcgctgga aacagtacag cgcctgctgc ctgtactgtg ccaggatcat
3061 ggactgacac ccgaacaggt ggtcgccatt gcttctaaca tcggaggacg gccagccttg
3121 gagtccatcg tagcccaatt gtccaggccc gatcccgcgt tggctgcgtt aacgaatgac
3181 catctggtgg cgttggcatg tcttggtgga cgacccgcgc tcgatgcagt caaaaagggt
3241 ctgcctcatg ctcccgcatt gatcaaaaga accaaccggc ggattcccga gagaacttcc
3301 catcgagtcg cgggatccca actagtcaaa agtgaactgg aggagaagaa atctgaactt
3361 cgtcataaat tgaaatatgt gcctcatgaa tatattgaat taattgaaat tgccagaaat
3421 tccactcagg atagaattct tgaaatgaag gtaatggaat tttttatgaa agtttatgga
3481 tatagaggta aacatttggg tggatcaagg aaaccggacg gagcaattta tactgtcgga
3541 tctcctattg attacggtgt gatcgtggat actaaagctt atagcggagg ttataatctg
3601 ccaattggcc aagcagatga aatgcaacga tatgtcgaag aaaatcaaac acgaaacaaa
3661 catatcaacc ctaatgaatg gtggaaagtc tatccatctt ctgtaacgga atttaagttt
3721 ttatttgtga gtggtcactt taaaggaaac tacaaagctc agcttacacg attaaatcat
3781 atcactaatt gtaatggagc tgttcttagt gtagaagagc ttttaattgg tggagaaatg
3841 attaaagccg gcacattaac cttagaggaa gtcagacgga aatttaataa cggcgagata
3901 aacttttaag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
3961 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
4021 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
4081 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
4141 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
4201 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
4261 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
4321 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
4381 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
4441 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
4501 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
4561 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
4621 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
4681 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
4741 tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc
4801 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
4861 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
4921 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
4981 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
5041 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
5101 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
5161 tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
5221 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
5281 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
5341 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
5401 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgtcgacagg
5461 tgcttctcga tctgcatcct gggatcaaag cgatagtgaa ggacagtgat ggacagccga
5521 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc
5581 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
5701 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
5881 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
6481 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
6721 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
7921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
7981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
8041 aaagtgccac ctgacgtc
//
