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ID PTKBETA preliminary; circular DNA; SYN; 7481 BP.
XX
AC IG1130; ATCC77178;
XX
DT 01-FEB-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE Vertebrate/E.coli plasmid vector pTKbeta - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RC pADbeta from adenovirus 2 MLP & SV40 splice/polyA & pUC19
RC pCMVbeta from CMV IEP & SV40 splice/polyA & pUC19
RC pSVbeta from SV40 early promoter & SV40 splice/polyA & pUC19
RC pTKbeta from HSV TK gene promoter & SV40 splice/polyA & pUC19
RA MacGregor G.R., Caskey C.T.;
RT "Construction of plasmids that express E. coli beta-galactosidase
RT in mammalian cells";
RL Nucleic Acids Res. 17:2365-2365(1989).
XX
RN [2]
RC pSV0cat, pSV2cat, pA10CAT2, pCAT..., pBLCAT2, pBLCAT3 from cat gene
RC pSK-SVCAT, pSK-CAT from cat gene
RC pRSVL-Adelta5', pSV2AL-Adelta5', pT81luc, pSluc2 from luciferase gene
RC pXP1, pXP2, pOLUC from luciferase gene
RC pXGH, pTKGH, pOGH from growth hormone gene
RC pSV2Apap, pSV232Apap, pSV0Apap from alkaline phosphatase gene
RC pBC12/RSV/SEAP, pBC12/PL/SEAP from secreted alkaline phosphatase
RC pCMVbeta, pSVbeta, pNASSbeta from beta-galactosidase gene
RA Alam J., Cook J.L.;
RT "Reporter genes: application to the study of mammalian gene
RT transcription";
RL Anal. Biochem. 188:245-254(1990).
XX
CC ATCC size is 7600 bp.
CC The order of the major features in this plasmid is: pUC19 - EcoRI -
CC HSV thymidine kinase promoter - SV40 late 16S/19S splice signals -
CC NotI - lacZ - NotI - SV40 late polyadenylation signal - XbaI -
CC HindIII - pUC19. [1]
CC The lacZ gene may be excised by NotI and replaced with alternative
CC sequences. [1]
CC Reporter plasmid permitting visual detection of beta-galactosidase
CC activity by histochemical staining. [1]
CC Designed to express low levels of beta-galactosidase or an
CC alternative gene product. May be used as a reference plasmid in
CC transfection experiments or to monitor the effect of trans-acting
CC factors. [1]
CC Restriction digest of the clone give the following sizes (kb):
CC EcoRI--7.2, 0.9; HindIII--7.6; EcoRI/HindIII--4.5,2.7,0.9;
CC NotI--4.4, 3.2; BamHI--3.8 (doublet). (ATCC staff)
CC Medium is -1 TB plus ampicillin (200 ug/ml).
CC NM (pTKbeta)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (CLONTECH)(ATCC)
CC HO (E.coli DH5alpha)(E.coli)(monkey COS cells)(vertebrate cells)
CC CP ()
CC FN (expression)(reporter)(promoter analysis)
CC SE ()
CC PA (pBR322)
CC BR (pADbeta)(pNASSbeta)(pCMVbeta)(pSVbeta)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pUC19 2686bp
FT 2. SV40, splice/polyA
FT -> plasmid
FT 1. plasmid NotI
FT 2. E. coli NotI-NotI, lacZ gene
FT -> plasmid2
FT 1. plasmid2
FT 2. HSV, tk promoter/#X14112
FT -> pTKbeta 7481bp"
FT misc_binding 0..0
FT /note="MCS EcoRI-BamHI-XbaI-SalI-PstI-SphI-HindIII"
FT misc_binding 0..0
FT /note="MCS unique XbaI-NotI-SalI-SphI-HindIII"
FT misc_binding 0..0
FT /note="SIT EcoRI"
FT misc_binding 0..0
FT /note="SIT SalI"
FT promoter 0..0
FT /note="PRO Herpes simplex virus (HSV)
FT thymidine kinase (TK) gene"
FT misc_binding 0..0
FT /note="SIT EcoRI"
FT promoter 0..0
FT /note="PRO SV40 enhancer"
FT misc_binding 0..0
FT /note="SIT BglII"
FT misc_binding 0..0
FT /note="SIT XhoI"
FT misc_feature 0..0
FT /note="SPL SV40 splicing sequences"
FT misc_binding 0..0
FT /note="SIT BamHI"
FT misc_binding 0..0
FT /note="SIT PstI"
FT misc_binding 0..0
FT /note="SIT unique NotI"
FT CDS 0..0
FT /note="GEN E. coli beta-galactosidase gene (lacZ')"
FT misc_binding 0..0
FT /note="SIT SacI"
FT misc_binding 0..0
FT /note="SIT NotI"
FT misc_binding 0..0
FT /note="SIT BamHI"
FT polyA_signal 0..0
FT /note="PLA SV40 early gene"
FT misc_binding 0..0
FT /note="SIT BamHI"
FT misc_binding 0..0
FT /note="SIT SalI"
FT misc_binding 0..0
FT /note="SIT PstI"
FT misc_binding 3900..3900
FT /note="SIT HindIII"
FT CDS complement(0..0)
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
XX
SQ Sequence 7481 BP; 1708 A; 1958 C; 2065 G; 1750 T; 0 other;
gaattcgtcg acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc
atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc
gcctgctgcg cgacggcgag ctgctcacca cccactcgac cagatccaaa tgagtcttcg
gacctcgcgg gggccgctta agcggtggtt agggtttgtc tgacgcgggg ggagggggaa
ggaacgaaac actctcattc ggaggcggct cggggtttgg tcttggtggc cacgggcacg
cagaagagcg ccgcgatcct cttaagcacc cccccgccct ccgtggaggt gggggtttgg
tcggcgggtg gtaactggcg ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac
cttttcggtc tgctcgcaga cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg
gtcctaggct caatggggac cgtatacgtg gacaggctct ggagcatccg cacgactgcg
gtgatattac cggagacctt ctgcgggacg agccgggtca cgcggctgac gcggagcgtc
cgttgggcga caaacaccag gacggggcac aggtacacta tcttgtcacc cggaggcgcg
agggactgca ggagcttcag ggagtggcgc agctgcttca tccccgtgac ccgttgccgc
gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct atgatgacac
aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag tcggggcggc
gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac accgagcgac
cctgcagcga cccgcttaac agcgtcaaca gcgtgccgca gatctgctcg acggtactcg
aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg
tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact
tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt acccgcggcc
gcaattcccg gggatcgaaa gagcctgcta aagcaaaaaa gaagtcacca tgtcgtttac
tttgaccaac aagaacgtga ttttcgttgc cggtctggga ggcattggtc tggacaccag
caaggagctg ctcaagcgcg atcccgtcgt tttacaacgt cgtgactggg aaaaccctgg
cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg
ttacggccag gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg
agaaaaccgc ctcgcggtga tggtgctgcg ttggagtgac ggcagttatc tggaagatca
ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac
acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact
ggaggctgaa gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt
atggcagggt gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga
tgagcgtggt ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact
gtggagcgcc gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg
cacgctgatt gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg
tctgctgctg ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca
tcctctgcat ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa
gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac
gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat
ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt
aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa
tgaatcaggc cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc
ttcccgcccg gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg
cccgatgtac gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat
caaaaaatgg ctttcgctac ctggagagac gcgcccgctg atcctttgcg aatacgccca
cgcgatgggt aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc
ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga
aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca
gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc
aaaacaccag cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag
cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg
taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat
tgaactgcct gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt
agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg
gcgtctggcg gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct
gaccaccagc gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg
ccagtcaggc tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct
gcgcgatcag ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg
cattgaccct aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc
agcgttgttg cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca
cgcgtggcag catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg
tagtggtcaa atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc
gcggattggc ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt
agggccgcaa gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct
gccattgtca gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg
gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag
ccgctacagt caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga
aggcacatgg ctgaatatcg acggtttcca tatggggatt ggtggcgacg actcctggag
cccgtcagta tcggcggaat tacagctgag cgccggtcgc taccattacc agttggtctg
gtgtcaaaaa taataataac cgggcaggcc atgtctgccc gtatttcgcg taaggaaatc
cattatgtac tatttaaaaa acacaaactt ttggatgttc ggtttattct ttttctttta
cttttttatc atgggagcct acttcccgtt tttcccgatt tggctacatg acatcaacca
tatcagcaaa agtgatacgg gtattatttt tgccgctatt tctctgttct cgctattatt
ccaaccgctg tttggtctgc tttctgacaa actcggcctc gactctaggc ggccgcgggg
atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
tgtgggaggt tttttcggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
cagggttttc ccagtcacga cgttgtaaaa cgacggccag t
//