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基本信息

質粒圖譜和多克隆位點信息

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

載體描述

EGFP were inserted into pcDNA3.1(+) by XhoI and XbaI，編號p1695,由轉導生物改造完成。注意，若目的基因插入MCS區域，目的基因需加起始密碼子。

載體序列

LOCUS       Exported                6148 bp ds-DNA     circular SYN 20-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pcDNA3.1?+?-EGFP

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6148)

  AUTHORS   BT-lab

  TITLE     Direct Submission

  JOURNAL   Exported Wednesday, Mar 20, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..6148

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        235..614

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        615..818

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early

                     promoter"

     primer_bind     829..848

                     /label=pcDNA3.1-F

     promoter        863..881

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    895..990

                     /label=5'MCS

     CDS             991..1710

                     /codon_start=1

                     /product="enhanced GFP"

                     /label=EGFP

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL

                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD

                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK

                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL

                     EFVTAAGITLGMDELYK"

     misc_feature    1711..1730

                     /label=3'MCS

     primer_bind     complement(1742..1759)

                     /label=pcDNA3.1-R (BGH-R)

     polyA_signal    1748..1972

                     /label=bGH poly(A) signal

                     /note="bovine growth hormone polyadenylation signal"

     rep_origin      2018..2446

                     /direction=RIGHT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     promoter        2460..2789

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2640..2775

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             2856..3650

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /label=NeoR/KanR

                     /note="confers resistance to neomycin, kanamycin, and G418

                     (Geneticin(R))"

                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     polyA_signal    3824..3945

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     primer_bind     complement(3994..4010)

                     /label=M13 rev

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

                     variants"

     promoter        complement(4042..4072)

                     /label=lac promoter

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

     rep_origin      complement(4396..4981)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(5152..6012)

                     /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"

     promoter        complement(6013..6117)

                     /gene="bla"

                     /label=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 gtctaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc

      961 agatatccag cacagtggcg gccgctcgag atggtgagca agggcgagga gctgttcacc

     1021 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg

     1081 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc

     1141 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag

     1201 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc

     1261 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc

     1321 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac

     1381 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac

     1441 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac

     1501 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc

     1561 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa

     1621 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc

     1681 actctcggca tggacgagct gtacaagtaa tctagagggc ccgtttaaac ccgctgatca

     1741 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc

     1801 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg

     1861 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg

     1921 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag

     1981 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta

     2041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg

     2101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa

     2161 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc

     2221 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt

     2281 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca

     2341 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc

     2401 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg

     2461 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca

     2521 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa

     2581 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca

     2641 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt

     2701 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag

     2761 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg

     2821 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg

     2881 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa

     2941 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg

     3001 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt

     3061 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa

     3121 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc

     3181 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg

     3241 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg

     3301 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg

     3361 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg

     3421 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact

     3481 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg

     3541 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc

     3601 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct

     3661 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac

     3721 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat

     3781 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc

     3841 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc

     3901 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc

     3961 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg

     4021 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg

     4081 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc

     4141 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt

     4201 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct

     4261 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga

     4321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc

     4381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg

     4441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg

     4501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt

     4561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt

     4621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg

     4681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact

     4741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt

     4801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct

     4861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac

     4921 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca

     4981 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta

     5041 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa

     5101 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg

     5161 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg

     5221 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc

     5281 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc

     5341 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa

     5401 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc

     5461 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg

     5521 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc

     5581 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat

     5641 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg

     5701 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc

     5761 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg

     5821 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat

     5881 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg

     5941 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg

     6001 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct

     6061 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac

     6121 atttccccga aaagtgccac ctgacgtc

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