| 质粒名称: | pHybrid-EBFP2 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0000205 |
| 产品用途: | Dual-expression vector. Expresses EBFP2 in bacterial cells and co-expresses EBFP2 with mScarlet in mammalian cells |
| 存储实验室: | KIRYL PIATKEVICH Lab |
|
载体骨架基本信息 |
|
| 载体名称: | pHybrid |
| 表达系统: | 哺乳动物表达、细菌表达 |
| 启动子: | rhaB, CMV, SV40 |
| 拷贝数: | 高拷贝 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a,TOP10 |
|
插入基因信息 |
|
| 插入基因名称: | EBFP2, mScarlet |
| 物种来源: | Synthetic, Synthetic |
| 密码子优化: | 哺乳细胞表达优化 |
|
文献引用方法 |
|
| 参考文献部分: |
Dual-expression system for blue fluorescent protein optimization
Papadaki S., Wang X., Wang Y., Zhang H., Jia S., Liu S., Yang M., Zhang D., Jia J., Köster R.W., Namikawa K., Piatkevich K.D.
Sci. Reports 2022 121 12, 1–16 (2022)
|
买家导航

pHybrid-EBFP2载体质粒基本信息
pHybrid-EBFP2质粒图谱载体图谱和pHybrid-EBFP2载体序列质粒序列多克隆位点信息

pHybrid-EBFP2质粒载体简介
pHybrid-EBFP2质粒序列载体序列
LOCUS pHybrid-EBFP2 5615 bp DNA circular SYN 31-MAY-2023
DEFINITION 72.Electra1TSN20211022-0571-00003_1023-ban2_H09_H09_i7.1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5615)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5615
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 754..858
/gene="bla"
/label=AmpR promoter
CDS 859..1719
/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"
rep_origin 1890..2478
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
enhancer 2689..3068
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3069..3272
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 3305..3423
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
misc_feature 3441..3446
/label=Shine-Dalgarno
CDS 3464..3481
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind 3487..3530
/label=1216-pCI-mBlueberry2-F
primer_bind 3487..3530
/label=1216-pCI-EBFP2/mBlueberry2-F
primer_bind 3487..3520
/label=1216-pCI-mTagBFP2-F
regulatory 3512..3521
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3518..4237
/codon_start=1
/product="enhanced blue variant of GFP (Ai et al., 2007)"
/label=EBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
EFRTAAGITLGMDELYK"
primer_bind complement(4214..4261)
/label=1216-pCI-mBlueberry2-R
primer_bind complement(4214..4261)
/label=1216-pCI-EBFP2/mBlueberry2-R
primer_bind complement(4235..4261)
/label=1216-pCI-mTagBFP2-R
terminator 4258..4344
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
polyA_signal 4368..4489
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4530..4740
/label=SV40 promoter
/note="SV40 early promoter"
promoter 4552..4748
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4599..4734
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4748..4767
/label=1216-pCI-mBlueberry2-F
primer_bind 4748..4767
/label=1216-pCI-EBFP2/mBlueberry2-F
regulatory 4749..4758
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4755..5450
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet-I
/note="matures faster than mScarlet"
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
primer_bind complement(5430..5451)
/label=1216-pCI-mBlueberry2-R
primer_bind complement(5430..5451)
/label=1216-pCI-EBFP2/mBlueberry2-R
polyA_signal 5455..5502
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin join(5582..5615,1..422)
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
61 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
181 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
301 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
361 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
421 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
481 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
541 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
601 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
661 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
721 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
841 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
1021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
1081 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
1141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
1201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
1261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
1321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
1381 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
1441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
1501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
1561 gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
1621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
1681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
1741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
1801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
1861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
1921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
1981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
2041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
2101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
2161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
2221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
2281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
2341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
2401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
2461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
2521 tggccttttg ctcacatggc tcgacagatc ttcaatattg gccattagcc atattattca
2581 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
2641 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
2701 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
2761 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
2821 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
2881 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
2941 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
3001 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
3061 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
3121 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
3181 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
3241 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
3301 gcttcaccac aattcagcaa attgtgaaca tcatcacgtt catctttccc tggttgccaa
3361 tggcccattt tcctgtcagt aacgagaagg tcgcgaattc aggcgctttt tagactggtc
3421 gtaacccgtt tttggctaac aggaggaatt aaccatgggg agccaccatc accatcacca
3481 tgctagcctc gagaattcac gcgtggtacc agccaccatg gtgagcaagg gcgaggagct
3541 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
3601 cagcgtgagg ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
3661 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg
3721 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
3781 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcacctacaa
3841 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
3901 cgtcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag
3961 ccacaacatc tatatcatgg ccgtcaagca gaagaacggc atcaaggtga acttcaagat
4021 ccgccacaac gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
4081 catcggcgac ggccccgtgc tgctgcccga cagccactac ctgagcaccc agtccgtgct
4141 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcc gcaccgccgc
4201 cgggatcact ctcggcatgg acgagctgta caagtaatct agagtcgacc caggcatcaa
4261 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
4321 aacgctctcc tgagtaggac aaatgcggcc gcttcgagca gacatgataa gatacattga
4381 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
4441 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
4501 ttgggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
4561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
4621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
4681 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
4741 tttgcaaagc caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca
4801 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
4861 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
4921 ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcatcaagc
4981 accccgccga catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc
5041 gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg
5101 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg
5161 taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg
5221 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg
5281 cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg
5341 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg
5401 aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag tgaccggcaa
5461 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcggatccgg gctggcgtaa
5521 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
5581 gacgcgccct gtagcggcgc attaagcgcg gcggg
//
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