LOCUS       SBE120_-_pJET1.2:(K):tN 3258 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(53..844)
                     /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(845..913)
                     /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(914..1018)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: bla"
     primer          1394..1413
                     /label="1125_PJET fw"
                     /note="sequence: CCATATCCATCCGGCGTAAT"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        1410..1428
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     primer          1476..1516
                     /label="203_K_T3_tNos_F"
                     /note="sequence: gcatGGTCTCagtatcgttcaaacatttggcaataaagttt"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1477..1478
                     /label="single point deletion"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="One C missing in between"
     misc_feature    1487..1490
                     /label="K gga_oh"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1491..1743
                     /label="nos terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(1723..1760)
                     /label="1304_S_T3_tNOS_R"
                     /note="sequence: TCTACCGGTCTCCccgaCCCGATCTAGTAACATAGATG"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1744..1747
                     /label="S gga_oh"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(1871..1890)
                     /label="1126_PJET rv"
                     /note="sequence: CCTGATGAGGTGGTTAGCAT"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     protein_bind    2171..2187
                     /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(2195..2201)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2202..2219)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2220..2225)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2552..3140)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
ORIGIN
        1 AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT
       61 GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT
      121 GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG
      181 CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG
      241 CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA
      301 ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG
      361 CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG
      421 GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT
      481 CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA
      541 TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG
      601 GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC
      661 CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG
      721 GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA
      781 TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG
      841 GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT
      901 GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC
      961 TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA
     1021 CATTTCCCCG AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG ACATTAACCT
     1081 ATAAAAATAG GCGTATCACG AGGCCGCCCC TGCAGCCGAA TTATATTATT TTTGCCAAAT
     1141 AATTTTTAAC AAAAGCTCTG AAGTCTTCTT CATTTAAATT CTTAGATGAT ACTTCATCTG
     1201 GAAAATTGTC CCAATTAGTA GCATCACGCT GTGAGTAAGT TCTAAACCAT TTTTTTATTG
     1261 TTGTATTATC TCTAATCTTA CTACTCGATG AGTTTTCGGT ATTATCTCTA TTTTTAACTT
     1321 GGAGCAGGTT CCATTCATTG TTTTTTTCAT CATAGTGAAT AAAATCAACT GCTTTAACAC
     1381 TTGTGCCTGA ACACCATATC CATCCGGCGT AATACGACTC ACTATAGGGA GAGCGGCCGC
     1441 CAGATCTTCC GGATGGCTCG AGTTTTTCAG CAAGATgatG GTCTCagtat cgttcaaaca
     1501 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     1561 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     1621 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     1681 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtCATCTATG TTACTAGATC
     1741 GGGtcggGGA GACCGGTAGA ATCTTTCTAG AAGATCTCCT ACAATATTCT CAGCTGCCAT
     1801 GGAAAATCGA TGTTCTTCTT TTATTCTCTC AAGATTTTCA GGCTGTATAT TAAAACTTAT
     1861 ATTAAGAACT ATGCTAACCA CCTCATCAGG AACCGTTGTA GGTGGCGTGG GTTTTCTTGG
     1921 CAATCGACTC TCATGAAAAC TACGAGCTAA ATATTCAATA TGTTCCTCTT GACCAACTTT
     1981 ATTCTGCATT TTTTTTGAAC GAGGTTTAGA GCAAGCTTCA GGAAACTGAG ACAGGAATTT
     2041 TATTAAAAAT TTAAATTTTG AAGAAAGTTC AGGGTTAATA GCATCCATTT TTTGCTTTGC
     2101 AAGTTCCTCA GCATTCTTAA CAAAAGACGT CTCTTTTGAC ATGTTTAAAG TTTAAACCTC
     2161 CTGTGTGAAA TTGTTATCCG CTCACAATTC CACACATTAT ACGAGCCGGA AGCATAAAGT
     2221 GTAAAGCCTG GGGTGCCTAA TGAGTGAGCT AACTCACATT AATTGCGTTG CGCTCACTGC
     2281 CAATTGCTTT CCAGTCGGGA AACCTGTCGT GCCAGCTGCA TTAATGAATC GGCCAACGCG
     2341 CGGGGAGAGG CGGTTTGCGT ATTGGGCGCT CTTCCGCTTC CTCGCTCACT GACTCGCTGC
     2401 GCTCGGTCGT TCGGCTGCGG CGAGCGGTAT CAGCTCACTC AAAGGCGGTA ATACGGTTAT
     2461 CCACAGAATC AGGGGATAAC GCAGGAAAGA ACATGTGAGC AAAAGGCCAG CAAAAGGCCA
     2521 GGAACCGTAA AAAGGCCGCG TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC
     2581 ATCACAAAAA TCGACGCTCA AGTCAGAGGT GGCGAAACCC GACAGGACTA TAAAGATACC
     2641 AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT TCCGACCCTG CCGCTTACCG
     2701 GATACCTGTC CGCCTTTCTC CCTTCGGGAA GCGTGGCGCT TTCTCATAGC TCACGCTGTA
     2761 GGTATCTCAG TTCGGTGTAG GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG
     2821 TTCAGCCCGA CCGCTGCGCC TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC
     2881 ACGACTTATC GCCACTGGCA GCAGCCACTG GTAACAGGAT TAGCAGAGCG AGGTATGTAG
     2941 GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC CTAACTACGG CTACACTAGA AGGACAGTAT
     3001 TTGGTATCTG CGCTCTGCTG AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT
     3061 CCGGCAAACA AACCACCGCT GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC
     3121 GCAGAAAAAA AGGATCTCAA GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT
     3181 GGAACGAAAA CTCACGTTAA GGGATTTTGG TCATGAGATT ATCAAAAAGG ATCTTCACCT
     3241 AGATCCTTTT AAATTAAA
//