pHybrid-EBFP2

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pHybrid-EBFP2载体质粒基本信息

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)

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载体图谱

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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