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ID   PCMVBETA   preliminary; circular DNA; SYN; 7164 BP.
XX
AC   IG1058; ATCC77177;
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 pCMVbeta - complete.
XX
KW   cloning vector.
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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 7300 bp.
CC   The order of the major features in this plasmid is [1]: pUC19 -
CC   EcoRI - CMV immediate early promoter/enhancer - SV40 late 16S/19S
CC   splice signals -NotI - lacZ - NotI - SV40 late polyadenylation
CC   signal - XbaI - HindIII - pUC19.
CC   The lacZ gene may be excised by NotI and replaced with alternative
CC   sequences. [1]
CC   Designed to express high 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   Reporter plasmid permitting visual detection of beta-galactosidase
CC   activity by histochemical staining. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--7.3; HindIII--7.3; EcoRI/HindIII--4.6, 2.7; NotI--3.8, 3.5;
CC   BamHI--3.65; 3.45; 0.2. (ATCC staff)
CC   Medium is -1 TB plus ampicillin (200 ug/ml).
CC   NM (pCMVbeta)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (CLONTECH)(ATCC)
CC   HO (E.coli DH5alpha)(monkey COS cells)(vertebrate cells)(E.coli)
CC   CP ()
CC   FN (expression)(reporter)(promoter analysis)
CC   SE ()
CC   PA ()
CC   BR (pADbeta)(pNASSbeta)(pSVbeta)(pTKbeta)
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. CMV, immediate early promoter
FT                   -> pCMVbeta 7164bp"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
FT   promoter        0..0
FT                   /note="PRO human cytomegalovirus (CMV)
FT                   immediate early gene"
FT   misc_binding    0..0
FT                   /note="SIT SacI"
FT   misc_binding    0..0
FT                   /note="SIT BamHI"
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 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)"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-BamHI-XbaI-SalI-PstI-SphI-HindIII"
XX
SQ   Sequence 7164 BP; 1700 A; 1840 C; 1897 G; 1727 T; 0 other;
     gaattcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
     tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     cgggaccgat ccagcctccg gactctagag gatccggtac tcgaggaact gaaaaaccag
     aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
     gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
     gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg gccgcaattc ccggggatcg
     aaagagcctg ctaaagcaaa aaagaagtca ccatgtcgtt tactttgacc aacaagaacg
     tgattttcgt tgccggtctg ggaggcattg gtctggacac cagcaaggag ctgctcaagc
     gcgatcccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac
     cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
     tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtaacctatc
     ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
     catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
     ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc caggacagtc
     gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac cgcctcgcgg
     tgatggtgct gcgttggagt gacggcagtt atctggaaga tcaggatatg tggcggatga
     gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc agcgatttcc
     atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct gaagttcaga
     tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag ggtgaaacgc
     aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt ggtggttatg
     ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc gccgaaatcc
     cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg attgaagcag
     aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg ctgctgaacg
     gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg catggtcagg
     tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac aactttaacg
     ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc gaccgctacg
     gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca atgaatcgtc
     tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga atggtgcagc
     gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca ggccacggcg
     ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc ccggtgcagt
     atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg tacgcgcgcg
     tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc
     tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg ggtaacagtc
     ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta cagggcggct
     tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc aacccgtggt
     cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt atgaacggtc
     tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac cagcagcagt
     ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc
     atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg
     gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac
     cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga
     ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg gcggaaaacc
     tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc agcgaaatgg
     atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca ggctttcttt
     cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat cagttcaccc
     gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac cctaacgcct
     gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca
     cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg
     ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga
     ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact
     gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact
     atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt
     ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc gaattgaatt
     atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac agtcaacagc
     aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca tggctgaata
     tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca gtatcggcgg
     aattacagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa aaataataat
     aaccgggcag gccatgtctg cccgtatttc gcgtaaggaa atccattatg tactatttaa
     aaaacacaaa cttttggatg ttcggtttat tctttttctt ttactttttt atcatgggag
     cctacttccc gtttttcccg atttggctac atgacatcaa ccatatcagc aaaagtgata
     cgggtattat ttttgccgct atttctctgt tctcgctatt attccaaccg ctgtttggtc
     tgctttctga caaactcggc ctcgactcta ggcggccgcg gggatccaga catgataaga
     tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttcg
     gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt
     cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     cgacgttgta aaacgacggc cagt
//