Return to this vector's summary.
ID 2MICRONA preliminary; circular DNA; FUN; 6318 BP.
XX
AC J01347; L00321; L00322; L00323; L00324; M10185; M11111; M11593;
AC M14239; M14240; M14241; M14242; M14243; M14244; M14245; M14253;
AC M14254; M14255; M14256; M14257; M14258; M14259; M14591; M14592;
AC M14593; M14594; M14595; M14596; M14597; M14598; V01323;
XX
DT 01-JUL-1994 (Rel. 9, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE Saccharomyces plasmid 2 micron A form - complete.
XX
KW cloning vector.
XX
OS Saccharomyces cerevisiae (yeast)
OC Eukaryota;; Fungi; Eumycota; Hemiascomycetes; Endomycetales;
OC Saccharomycetaceae.
XX
RN [1]
RP 1-1022
RC pJDB71 from pMB9 & yeast 2 micron form A
RA Hindley J., Phear G.A.;
RT "Sequence of 1019 nucleotides encompassing one of the inverted
RT repeats from the yeast 2 micron plasmid";
RL Nucleic Acids Res. 7:361-375(1979).
XX
RN [2]
RP 1-6318
RC p82-6B from pMB9 & yeast 2 micron
RA Hartley J.L., Donelson J.E.;
RT "Nucleotide sequence of the yeast plasmid";
RL Nature 286:860-865(1980).
XX
RN [3]
RP 3891-3990
RC from yeast 2 micron, recombination site
RA Broach J.R., Guarascio V.R., Jayaram M.;
RT "Recombination within the yeast plasmid 2-micron circle is
RT site-specific";
RL Cell 29:227-234(1982).
XX
RN [4]
RP 3881-4020
RC pMMD2 from pBR322 & yeast LEU2 gene & yeast URA3 gene & yeast 2 micron
RA McLeod M., Volkert F., Broach J.R.;
RT "Components of the site-specific recombination system encoded by the
RT yeast plasmid 2-micron circle";
RL Cold Spring Harb. Symp. Quant. Biol. 49:779-787(1984).
XX
RN [5]
RP 670-732
RC pBA112 from pUC9 & yeast FLP recombination site
RA Andrews B.J., Proteau G.A., Beatty L.G., Sadowski P.D.;
RT "The FLP recombinase of the 2 micron circle DNA of yeast:
RT interaction with its target sequences";
RL Cell 40:795-803(1985).
XX
RN [6]
RP 5570-5605
RC plasmid from yeast 2 micron FLP & pMA301
RC pDV64 from plasmid & pDR540
RC pBA101 from pCV19 & pBR322
RC pBA104 from pBA101 & pCV19
RC pBA107 from pBA101 & pCV19
RA Babineau D., Vetter D., Andrews B.J., Gronostajski R.M.,
RA Proteau G.A., Beatty L.G., Sadowski P.D.;
RT "The FLP protein of the 2-micron plasmid of yeast: purification of
RT the protein from Escherichia coli cells expressing the cloned FLP
RT gene";
RL J. Biol. Chem. 260:12313-12319(1985).
XX
RN [7]
RC plasmid from pUC9 & yeast 2 micron, FLP cleavage site
RC pGP40 from plasmid & yeast 2 micron
RA Gronostajski R.M., Sadowski P.D.;
RT "Determination of DNA sequences essential for FLP-mediated
RT recombination by a novel method";
RL J. Biol. Chem. 260:12320-12327(1985).
XX
RN [8]
RC pBR2micronA from pBR322 & yeast 2 micron A form
RC from yeast 2 micron, CAP site/polyA
RA Sutton A., Broach J.R.;
RT "Signals for transcription initiation and termination in the
RT Saccharomyces cerevisiae plasmid 2 micron circle";
RL Mol. Cell. Biol. 5:2770-2780(1985).
XX
RN [9]
RC from pBA112, FLP recombination site
RA Gronostajski R.M., Sadowski P.D.;
RT "The FLP recombinase of the Saccharomyces cerevisiae 2-micron
RT plasmid attaches covalently to DNA via a phosphotyrosyl linkage";
RL Mol. Cell. Biol. 5:3274-3279(1985).
XX
RN [10]
RP 667-739
RC pJFS166 from pXf3 & linker & yeast 2 micron, FLP binding site
RC pJFS122, pJFS110, pJFS105, pJFS100 from pJFS166 & M13mp8
RC pJFS30 series from pJFS100
RA Senecoff J.F., Bruckner R.C., Cox M.M.;
RT "The FLP recombinase of the yeast 2-micron-m plasmid:
RT Characterization of its recombination site";
RL Proc. Natl. Acad. Sci. U.S.A. 82:7270-7274(1985).
XX
RN [11]
RC pWP series from pMMD2 & linker
RC pMu5T2-2, p545-2, p624B-2, p613B-2 from pMMD2
RC pINV1 from pMMD2 & pCV20
RA McLeod M., Craft S., Broach J.R.;
RT "Identification of the crossover site during FLP-mediated
RT recombination in the Saccharomyces cerevisiae plasmid 2 micron
RT circle";
RL Mol. Cell. Biol. 6:3357-3367(1986).
XX
RN [12]
RP 1-6318
RC yeast 2 micron
RA Senecoff J.;
RT ;
RL Submitted (24-JAN-1986) by:
RL Senecoff J., .
XX
RN [13]
RC from yeast 2 micron
RC from yeast 2 micron & pBR322 & LEU2 gene
RA Broach J.R., Hicks J.B.;
RT "Replication and recombination functions associated with the yeast
RT 2micron circle";
RL Cell 21:501-508(1980).
XX
RN [14]
RC from yeast 2 micron
RA Broach J.R.;
RT "The yeast plasmid 2 micron circle";
RL Cell 28:203-204(1982).
XX
RN [15]
RC pMA301 from YEp series
RA Kingsman S., Kingsman A.;
RT ;
RL Unpublished (1985).
XX
RN [16]
RC pCV20 or pMJ5 from pBR322 & yeast 2 micron & LEU2 gene
RC pXho series from pCV20
RA Jayaram M., Li Y.Y., Broach J.R.;
RT "The yeast plasmid 2 micron circle encodes components required for
RT its high copy propagation";
RL Cell 34:95-104(1983).
XX
CC Yeast 2 micron plasmid contains two 599 bp inverted repeats
CC separated by a large unique (UL) and a small unique (US) region.
CC During recombination the UL and US regions invert producing two
CC sequence forms that differ in the orientation of one unique region
CC relative to the other. The A form is presented below.
CC FLP is the only 2-micron circle-encoded protein needed for specific
CC site recombination between the IRs of 2-micron circle. The minimal
CC size of the recombination site required for efficient FLP
CC recombinase-catalyzed recombination in vitro is no more than 28 bp,
CC which includes parts of two 13 bp inverted repeats (positions
CC 690-702 and 711-723) and all of an 8 bp spacer (703-710) [5]. The
CC FLP recombinase cleaves the DNA at the boundaries of the spacer and
CC becomes covalently linked to the spacer DNA [5],[9]. The
CC efficiency of the recombination is reduced if the spacer in a
CC recombinant site is increased or decreased by 1 bp, while the
CC spacer in the second site is unaltered [5]. Recombination between
CC two sites with identical 1-base pair additions or deletions is
CC relatively unaffected, suggesting that pairing of sequences in the
CC spacer regions is important in FLP-promoted recombination events
CC [5]. The sequence asymmetry utilized by the recombinase to
CC determine the orientation of the site is located uniquely within
CC the spacer region. Another 13 bp direct repeat, is found at
CC positions 676-688 [5]. FLP-mediated recombination involving two
CC FLP sites that are inverted with respect to each other results in
CC inversion of the DNA sequences between the sites [4]. If the
CC participating recombination sites are in direct orientation, FLP
CC promotes only the excision of the intervening DNA sequences [4].
CC The Rep 1 and Rep proteins are involved plasmid partitioning and
CC protein stability.
CC A start codon in phase with the Rep1 coding region is located at
CC positions 1966-1964. Two CAP sites for Rep1 mRNA are located
CC beyond the 'atg' codon (position 2008) at positions 2004 and 2005.
CC Complete source information:
CC Yeast (S.cerevisiae, strain A364A D5) DNA, clones pJDB71 [1],
CC p82-6B [2], pCV20 [3], pMMD2 [4], pGP20 [5], pJFS166 [10].
CC NCBI gi: 172190 & 4182
CC NM (yeast 2 micron A form)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP ()
CC HO (S.cerevisiae)
CC CP (high)
CC FN (cloning)
CC SE ()
CC PA (2 micron)
CC BR ()
CC OF (YEp)(YIp)(2micron Circle-pBR322-LEU2)
CC OR (1 bp upstream of EcoRI site)
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. S. cerevisiae
FT -> yeast 2 micron 6318bp"
FT mRNA join(5549..6318,1..545)
FT /note="Flp mRNA"
FT repeat_region 341..939
FT /note="IR2"
FT misc_binding 673..722
FT /note="FLP recombinase binding site A [9]"
FT misc_recomb 702..703
FT /note="US end/UL start [9],[7]"
FT mRNA complement(836..2004)
FT /note="Rep1 mRNA (alt.)"
FT mRNA complement(836..2005)
FT /note="Rep1 mRNA (alt.)"
FT mRNA complement(836..2010)
FT /note="Rep1 mRNA (alt.)"
FT mRNA complement(836..2017)
FT /note="Rep1 mRNA (alt.)"
FT mRNA complement(836..2019)
FT /note="Rep1 mRNA (alt.)"
FT mRNA complement(836..2038)
FT /note="Rep1 mRNA (alt.)"
FT mRNA 2254..2841
FT /note="D mRNA (alt.; 5' end +/- 3 bp)"
FT mRNA 2254..2861
FT /note="D mRNA (alt.; 5' end +/- 3 bp)"
FT repeat_region 3714..4312
FT /note="IR1"
FT misc_binding 3930..3979
FT /note="FLP-recombinase binding site B [9]"
FT misc_recomb 3941..3942
FT /note="UL end/US start [9],[7]"
FT mRNA complement(4108..5184)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5183)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5182)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5198)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5197)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5196)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5195)
FT /note="REP2 mRNA (major alt.)"
FT mRNA complement(4108..5212)
FT /note="REP2 mRNA (minor alt.)"
FT mRNA complement(4108..5223)
FT /note="REP2 mRNA (minor alt.)"
FT CDS join(5570..6318,1..523)
FT /note="recombinase; (FLP); NCBI gi: 172191."
FT CDS complement(887..2008)
FT /note="Rep 1 protein; NCBI gi: 172192."
FT CDS 2271..2816
FT /note="D protein; NCBI gi: 172193."
FT CDS complement(4308..5198)
FT /note="Rep 2 protein; NCBI gi: 172194."
XX
SQ Sequence 6318 BP; 1876 A; 1284 C; 1179 G; 1979 T; 0 other;
gaattctgaa ccagtcctaa aacgagtaaa taggaccggc aattcttcaa gcaataaaca
ggaataccaa ttattaaaag ataacttagt cagatcgtac aataaagctt tgaagaaaaa
tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa tctcacattg gaagacattt
gatgacctca tttctttcaa tgaagggcct aacggagttg actaatgttg tgggaaattg
gagcgataag cgtgcttctg ccgtggccag gacaacgtat actcatcaga taacagcaat
acctgatcac tacttcgcac tagtttctcg gtactatgca tatgatccaa tatcaaagga
aatgatagca ttgaaggatg agactaatcc aattgaggag tggcagcata tagaacagct
aaagggtagt gctgaaggaa gcatacgata ccccgcatgg aatgggataa tatcacagga
ggtactagac tacctttcat cctacataaa tagacgcata taagtacgca tttaagcata
aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg cagatatagg
tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg aagcgctcgt
tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac
ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
ttcgctcctt gaacttgcat ctaaactcga cctctacatc aacaggcttc caatgctctt
caaattttac tgtcaagtag acccatacgg ctgtaatatg ctgctcttca taatgtaagc
ttatctttat cgaatcgtgt gaaaaactac taccgcgata aacctttacg gttccctgag
attgaattag ttcctttagt atatgataca agacactttt gaactttgta cgacgaattt
tgaggttcgc catcctctgg ctatttccaa ttatcctgtc ggctattatc tccgcctcag
tttgatcttc cgcttcagac tgccattttt cacataatga atctatttca ccccacaatc
cttcatccgc ctccgcatct tgttccgtta aactattgac ttcatgttgt acattgttta
gttcacgaga agggtcctct tcaggcggta gctcctgatc tcctatatga cctttatcct
gttctctttc cacaaactta gaaatgtatt catgaattat ggagcaccta ataacattct
tcaaggcgga gaagtttggg ccagatgccc aatatgcttg acatgaaaac gtgagaatga
atttagtatt attgtgatat tctgaggcaa ttttattata atctcgaaga taagagaaga
atgcagtgac ctttgtattg acaaatggag attccatgta tctaaaaaat acgcctttag
gccttctgat accctttccc ctgcggttta gcgtgccttt tacattaata tctaaaccct
ctccgatggt ggcctttaac tgactaataa atgcaaccga tataaactgt gataattctg
ggtgatttat gattcgatcg acaattgtat tgtacactag tgcaggatca ggccaatcca
gttctttttc aattaccggt gtgtcgtctg tattcagtac atgtccaaca aatgcaaatg
ctaacgtttt gtatttctta taattgtcag gaactggaaa agtccccctt gtcgtctcga
ttacacacct actttcatcg tacaccatag gttggaagtg ctgcataata cattgcttaa
tacaagcaag cagtctctcg ccattcatat ttcagttatt ttccattaca gctgatgtca
ttgtatatca gcgctgtaaa aatctatctg ttacagaagg ttttcgcggt ttttataaac
aaaactttcg ttacgaaatc gagcaatcac cccagctgcg tatttggaaa ttcgggaaaa
agtagagcaa cgcgagttgc attttttaca ccataatgca tgattaactt cgagaaggga
ttaaggctaa tttcactagt atgtttcaaa aacctcaatc tgtccattga atgccttata
aaacagctat agattgcata gaagagttag ctactcaatg ctttttgtca aagcttactg
atgatgatgt gtctactttc aggcgggtct gtagtaagga gaatgacatt ataaagctgg
cacttagaat tccacggact atagactata ctagtatact ccgtctactg tacgatacac
ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc
cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta
gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat
ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta
atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg
aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat
atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg
catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc
ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa
aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc
agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac
ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca taccctagaa
gtattacgtg attttctgcc ccttaccctc gttgctactc tccttttttt cgtgggaacc
gctttagggc cctcagtgat ggtgttttgt aatttatatg ctcctcttgc atttgtgtct
ctacttcttg ttcgcctgga gggaacttct tcatttgtat tagcatggtt cacttcagtc
cttccttcca actcactctt tttttgctgt aaacgattct ctgccgccag ttcattgaaa
ctattgaata tatcctttag agattccggg atgaataaat cacctattaa agcagcttga
cgatctggtg gaactaaagt aagcaattgg gtaacgacgc ttacgagctt cataacatct
tcttccgttg gagctggtgg gactaataac tgtgtacaat ccatttttct catgagcatt
tcggtagctc tcttcttgtc tttctcgggc aatcttccta ttattatagc aatagatttg
tatagttgct ttctattgtc taacagcttg ttattctgta gcatcaaatc tatggcagcc
tgacttgctt cttgtgaaga gagcatacca tttccaatcg aatcaaacct ttccttaacc
atcttcgcag caggcaaaat tacctcagca ctggagtcag aagatacgct ggaatcttct
gcgctagaat caagaccata cggcctaccg gttgtgagag attccatggg ccttatgaca
tatcctggaa agagtagctc atcagactta cgtttactct ctatatcaat atctacatca
ggagcaatca tttcaataaa cagccgacat acatcccaga cgctataagc tgtacgtgct
tttaccgtca gattcttggc tgtttcaatg tcgtccattt tggttttctt ttaccagtat
tgttcgtttg ataatgtatt cttgcttatt acattataaa atctgtgcag atcacatgtc
aaaacaactt tttatcacaa gatagtaccg caaaacgaac ctgcgggccg tctaaaaatt
aaggaaaagc agcaaaggtg catttttaaa atatgaaatg aagataccgc agtaccaatt
attttcgcag tacaaataat gcgcggccgg tgcatttttc gaaagaacgc gagacaaaca
ggacaattaa agttagtttt tcgagttagc gtgtttgaat actgcaagat acaagataaa
tagagtagtt gaaactagat atcaattgca cacaagatcg gcgctaagca tgccacaatt
tggtatatta tgtaaaacac cacctaaggt gcttgttcgt cagtttgtgg aaaggtttga
aagaccttca ggtgagaaaa tagcattatg tgctgctgaa ctaacctatt tatgttggat
gattacacat aacggaacag caatcaagag agccacattc atgagctata atactatcat
aagcaattcg ctgagtttcg atattgtcaa taaatcactc cagtttaaat acaagacgca
aaaagcaaca attctggaag cctcattaaa gaaattgatt cctgcttggg aatttacaat
tattccttac tatggacaaa aacatcaatc tgatatcact gatattgtaa gtagtttgca
attacagttc gaatcatcgg aagaagcaga taagggaaat agccacagta aaaaaatgct
taaagcactt ctaagtgagg gtgaaagcat ctgggagatc actgagaaaa tactaaattc
gtttgagtat acttcgagat ttacaaaaac aaaaacttta taccaattcc tcttcctagc
tactttcatc aattgtggaa gattcagcga tattaagaac gttgatccga aatcatttaa
attagtccaa aataagtatc tgggagtaat aatccagtgt ttagtgacag agacaaagac
aagcgttagt aggcacatat acttctttag cgcaaggggt aggatcgatc cacttgtata
tttggatgaa tttttgag
//