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ID   PNASSBETA  preliminary; circular DNA; SYN; 6541 BP.
XX
AC   IG1095; ATCC77180;
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 pNASSbeta - 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).
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CC   ATCC size is 6700 bp.
CC   The order of the major features in this plasmid is: pUC19 - EcoRI -
CC   SV40 late 16S/19S splice signals - NotI - lacZ - NotI - SV40 late
CC   polyadenylation signal - XbaI - HindIII - pUC19. [1]
CC   The lacZ gene may be excised by NotI and replaced with alternative
CC   sequences. [1]
CC   Promoter-less reporter plasmid permitting visual detection of
CC   transcription/translation activity by histochemical staining. [1]
CC   Promoters may be introduced upstream of the lacZ cassette at the
CC   unique EcoRI site. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.7; HindIII--6.7; EcoRI/HindIII--4.0, 2.7; NotI--3.5, 3.2;
CC   BamHI--3.65, 2.85, 0.2. (ATCC staff)
CC   Medium is -1 TB plus ampicillin (200 ug/ml).
CC   NM (pNASSbeta)
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)(pCMVbeta)(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                   -> pNASSbeta 6541bp"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT SacI"
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    200..200
FT                   /note="SIT NotI"
FT   CDS             200..3700
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ)"
FT   misc_binding    0..0
FT                   /note="SIT SacI"
FT   misc_binding    3700..3700
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 XbaI"
FT   misc_binding    0..0
FT                   /note="SIT SalI"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
FT   misc_binding    0..0
FT                   /note="SIT SphI"
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 6541 BP; 1549 A; 1675 C; 1744 G; 1573 T; 0 other;
     gaattcgagc tcggtactcg 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
//