The pGEM?-11Zf(?) Vector(a) can be used as a standard cloning vector, as a template
for in vitro transcription and for the production of ssDNA. The plasmid contains T7 and
SP6 RNA polymerase promoters flanking a multiple cloning region within the α-peptide
coding region of β-galactosidase (1). Insertional inactivation of the α-peptide allows
recombinant clones to be directly identified by color screening on indicator plates when
using appropriate E. coli strains (e.g., JM109, DH5α?, XL1-Blue). The multiple cloning
region contains unique restriction sites for Sfi I, Sac I, EcoR I, Sal I, Xho I, BamH I,
Apa I, Xba I, Not I, Sph I, Nsi I and Hind III.
For induction of ssDNA, bacterial cells with the F′ episome (e.g., JM109, XL1-Blue,
DH5α?F′) containing pGEM?-11Zf(?) recombinants are infected with an appropriate
helper phage. The plasmid then enters the f1 replication mode, and the resulting
ssDNA is exported from the cell as an encapsidated virus-like particle. The sequence of
the ssDNA rescued upon infection with helper phage is the same strand as that shown
in Figure 1. The exported ssDNA can be used for in vitro mutagenesis or can be
sequenced using Promega's SP6 Promoter Primer or pUC/M13 Reverse Primer.
The sequences of Promega vectors are available online at www.promega.com/vectors/
and are also available from the GenBank? database.
LOCUS pGEM-11Zf- 3223 bp DNA SYN
DEFINITION pGEM-11Zf-
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..3223
/organism="pGEM-11Zf-"
/mol_type="other DNA"
misc_feature complement(93..110)
/label="Sp6_primer"
promoter complement(124..142)
/label="M13_reverse_primer"
misc_feature complement(141..163)
/label="M13_pUC_rev_primer"
promoter complement(177..206)
/label="lac_promoter"
rep_origin complement(515..1134)
/label="pBR322_origin"
gene complement(1289..2149)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(1289..2149)
/label="ORF frame 1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(2191..2219)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature complement(2378..2400)
/label="pGEX_3_primer"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
rep_origin 2605..2911
/label="f1_origin"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature complement(3055..3203)
/label="lacZ_a"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature 3169..3191
/label="M13_pUC_fwd_primer"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter 3184..3200
/label="M13_forward20_primer"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter 3207..2
/label="T7_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 gggcgaattg gccaagtcgg ccgagctcga attcgtcgac ctcgagggat ccgggccctc
61 tagatgcggc cgcatgcata agcttgagta ttctatagtg tcacctaaat agcttggcgt
121 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
181 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
241 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttcgataggc
541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
961 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttggcat tgctacaggc atcgtggtgt
1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
1861 gagaataccg cgcccggcga ccgagttgct cttgcccggc gtcaatacgg gataatagtg
1921 tatgacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
2281 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
2341 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
2401 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
2461 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
2521 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
2581 tcaggcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
2641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
2701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
2761 cgatttagag ctttacggca cctcgaccgc aaaaaacttg atttgggtga tggttcacgt
2821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
2881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
2941 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
3001 atatttaacg cgaattttaa caaaatatta acgtttacaa tttccattcg ccattcaggc
3061 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
3121 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
3181 gttgtaaaac gacggccagt gaattgtaat acgactcact ata
//