LOCUS       SBE118_-_pJET1.2:(E):tN 3257 bp ds-DNA     circular     22-NOV-2022
DEFINITION  .
KEYWORDS    "accession:EF694056" "creator:Thermo Fisher (Fermentas)"
            "marker:AmpR" "organism:Escherichia coli"
COMMENT     Positive selection cloning vector with a lethal insert that allows
            for efficient recovery of blunt-ended PCR products. Linearize at
            Eco32I (EcoRV) to insert a blunt PCR product.
FEATURES             Location/Qualifiers
     CDS             complement(38..829)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="codon_start: 1 gene: bla note: confers resistance to ampicillin, carbenicillin, and related antibiotics product: β-lactamase transl_table: 1 translation: MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     CDS             complement(830..898)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="codon_start: 1 gene: bla note: confers resistance to ampicillin, carbenicillin, and related antibiotics product: β-lactamase transl_table: 1 translation: MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(899..1003)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: bla"
     primer          1379..1398
                     /label="1125_PJET fw"
                     /note="sequence: CCATATCCATCCGGCGTAAT"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        1395..1413
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     primer          1462..1500
                     /label="200_F_T1_tNos_R"
                     /note="sequence: atgcGGTCTCaaagccccgatctagtaacatagatgaca"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1473..1476
                     /label="F gga_oh"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(1477..1729)
                     /label="nos terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(1703..1744)
                     /label="199_E_T1_tNos_F"
                     /note="sequence: gcatGGTCTCatctacgttcaaacatttggcaataaagtttc"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1730..1733
                     /label="E gga_oh"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(1855..1874)
                     /label="1126_PJET rv"
                     /note="sequence: CCTGATGAGGTGGTTAGCAT"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     protein_bind    2155..2171
                     /label="lac operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
                     /note="bound_moiety: lac repressor encoded by lacI 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-β-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2179..2185)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2186..2203)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2204..2209)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2536..3124)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
ORIGIN
        1 CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG
       61 CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT
      121 AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG
      181 ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC
      241 GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG
      301 CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA
      361 TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA
      421 GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA
      481 TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA
      541 ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA
      601 AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG
      661 ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG
      721 GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG
      781 CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG
      841 GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC
      901 TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA
      961 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG
     1021 TGCCACCTGA CGTCTAAGAA ACCATTATTA TCATGACATT AACCTATAAA AATAGGCGTA
     1081 TCACGAGGCC GCCCCTGCAG CCGAATTATA TTATTTTTGC CAAATAATTT TTAACAAAAG
     1141 CTCTGAAGTC TTCTTCATTT AAATTCTTAG ATGATACTTC ATCTGGAAAA TTGTCCCAAT
     1201 TAGTAGCATC ACGCTGTGAG TAAGTTCTAA ACCATTTTTT TATTGTTGTA TTATCTCTAA
     1261 TCTTACTACT CGATGAGTTT TCGGTATTAT CTCTATTTTT AACTTGGAGC AGGTTCCATT
     1321 CATTGTTTTT TTCATCATAG TGAATAAAAT CAACTGCTTT AACACTTGTG CCTGAACACC
     1381 ATATCCATCC GGCGTAATAC GACTCACTAT AGGGAGAGCG GCCGCCAGAT CTTCCGGATG
     1441 GCTCGAGTTT TTCAGCAAGA TatgcGGTCT Caaagccccg atctagtaac atagatgaca
     1501 ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa
     1561 tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca
     1621 tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac
     1681 cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacgt agatGAGACC
     1741 atgcATCTTT CTAGAAGATC TCCTACAATA TTCTCAGCTG CCATGGAAAA TCGATGTTCT
     1801 TCTTTTATTC TCTCAAGATT TTCAGGCTGT ATATTAAAAC TTATATTAAG AACTATGCTA
     1861 ACCACCTCAT CAGGAACCGT TGTAGGTGGC GTGGGTTTTC TTGGCAATCG ACTCTCATGA
     1921 AAACTACGAG CTAAATATTC AATATGTTCC TCTTGACCAA CTTTATTCTG CATTTTTTTT
     1981 GAACGAGGTT TAGAGCAAGC TTCAGGAAAC TGAGACAGGA ATTTTATTAA AAATTTAAAT
     2041 TTTGAAGAAA GTTCAGGGTT AATAGCATCC ATTTTTTGCT TTGCAAGTTC CTCAGCATTC
     2101 TTAACAAAAG ACGTCTCTTT TGACATGTTT AAAGTTTAAA CCTCCTGTGT GAAATTGTTA
     2161 TCCGCTCACA ATTCCACACA TTATACGAGC CGGAAGCATA AAGTGTAAAG CCTGGGGTGC
     2221 CTAATGAGTG AGCTAACTCA CATTAATTGC GTTGCGCTCA CTGCCAATTG CTTTCCAGTC
     2281 GGGAAACCTG TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT
     2341 GCGTATTGGG CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT
     2401 GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG AATCAGGGGA
     2461 TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG GCCAGGAACC GTAAAAAGGC
     2521 CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC GAGCATCACA AAAATCGACG
     2581 CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG
     2641 AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT
     2701 TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC TCAGTTCGGT
     2761 GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC CCCGTTCAGC CCGACCGCTG
     2821 CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA AGACACGACT TATCGCCACT
     2881 GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT
     2941 CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGGACA GTATTTGGTA TCTGCGCTCT
     3001 GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA AACAAACCAC
     3061 CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT ACGCGCAGAA AAAAAGGATC
     3121 TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT CAGTGGAACG AAAACTCACG
     3181 TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA
     3241 AAAATGAAGT TTTAAAT
//