LOCUS       Exported                3103 bp ds-DNA    circular SYN 21-1-2014
DEFINITION  Multi-purpose cloning vector with T7 and SP6 promoters, and with a 
            multiple cloning site that encodes a cleavable N-terminal S-tag.
ACCESSION   .
VERSION     .
KEYWORDS    pT7Blue-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3103)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3103
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     CDS             1..549
                     /codon_start=1
                     /gene="lacZ (truncated)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="LacZ-alpha"
                     /translation="MTMITPSSNTTHYRESSLVLFAEAQNKRSTLATMGVTKETAAAKF
                     ERQHIDSGTGGSDDDDKSPGFSSKLQTRYVSDPYLGDTIEFRYLVDKLLEPRLALDHTC
                     GGPSSRPHNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLN
                     GEWEIVNVNILLKFALNFC"
     promoter        24..42
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5UTR           52..94
                     /note="Xenopus globin 5-UTR"
                     /note="translational enhancer from the 5-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    92..334
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             100..102
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             112..156
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHIDS"
     CDS             172..186
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     promoter        236..254
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(342..358)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(500..955)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        982..1086
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1087..1947
                     /codon_start=1
                     /note="AmpR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2118..2706
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3030..3060
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3068..3084
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3092..5
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"