LOCUS       SBE114_-_pJET1.2:(I):pA 3730 bp ds-DNA     circular     18-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(16..1509)
                     /label="Eco47I/T7"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
                     /note="codon_start: 1 function: gene: eco47IR note: cleaves double-stranded DNA at GGWCC sites in the absence of protective methylation product: mutant version of the Eco47I restriction endonuclease protein_id: transl_table: 1 translation: MSKETSFVKNAEELAKQKMDAINPELSSKFKFLIKFLSQFPEACSKPRSKKMQNKVGQEEHIEYLARSFHESRLPRKPTPPTTVPDEVVSIVLNISFNIQPENLERIKEEHRFSMAAENIVGDLLERYLAEKLEPSGRSGGRSPYSESYYAGWIWCSGTSVKAVDFIHYDEKNNEWNLLQVKNRDNTENSSSSKIRDNTTIKKWFRTYSQRDATNWDNFPDEVSSKNLNEEDFRAFVKNYLAKII*"
     primer          289..308
                     /label="1125_PJET fw"
                     /note="sequence: CCATATCCATCCGGCGTAAT"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        305..323
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     misc_feature    328..1178
                     /label="MCS"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="note: multiple cloning site with a central Eco32I (EcoRV) site"
     primer          372..410
                     /label="198_J_P3_pADH2_R"
                     /note="sequence: atgcGGTCTCagtggtgtgactgtcggagacgtggcagc"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    383..386
                     /label="J gga_oh"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(387..410)
                     /label="pADH2_R_Sun"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    complement(387..1111)
                     /label="pADH2_Sun"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    559..559
                     /label="point mutation"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="original nucleotide is C"
     misc_feature    complement(1093..1111)
                     /label="pADH2_F_Sun"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(1093..1127)
                     /label="197_I_P3_pADH2_F"
                     /note="sequence: gcatGGTCTCagtcacggctgaggcttccccgacg"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     misc_feature    1113..1116
                     /label="I gga_oh"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(1238..1257)
                     /label="1126_PJET rv"
                     /note="sequence: CCTGATGAGGTGGTTAGCAT"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     protein_bind    1538..1554
                     /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(1562..1568)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(1569..1586)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(1587..1592)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(1919..2507)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(2678..3469)
                     /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(3470..3538)
                     /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(3539..3643)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="gene: bla"
ORIGIN
        1 GCCCCTGCAG CCGAATTATA TTATTTTTGC CAAATAATTT TTAACAAAAG CTCTGAAGTC
       61 TTCTTCATTT AAATTCTTAG ATGATACTTC ATCTGGAAAA TTGTCCCAAT TAGTAGCATC
      121 ACGCTGTGAG TAAGTTCTAA ACCATTTTTT TATTGTTGTA TTATCTCTAA TCTTACTACT
      181 CGATGAGTTT TCGGTATTAT CTCTATTTTT AACTTGGAGC AGGTTCCATT CATTGTTTTT
      241 TTCATCATAG TGAATAAAAT CAACTGCTTT AACACTTGTG CCTGAACACC ATATCCATCC
      301 GGCGTAATAC GACTCACTAT AGGGAGAGCG GCCGCCAGAT CTTCCGGATG GCTCGAGTTT
      361 TTCAGCAAGA TatgcGGTCT Cagtggtgtg actgtcggag acgtggcagc tgtcagctag
      421 acgggaaatg acggtgcgag agtgaacagc cgggaatggg aacgcacgaa gagagagggc
      481 tgcgaaggct ggaccggtat gggctgagtg cgagtgctgt gcgagggtgt gtagcagtgt
      541 gtgcaagtga agagcaagTt gtaagagaga gagcgagcgc aaggttgcgg gacggtcgat
      601 gggtggggaa ggcgaagagc gaggaaggcc actcctgctc gggacgactc gcaagtcggc
      661 tcgctctcgc acgagacacg cttaattgac gcccacagcc actcgagcac gggacccagc
      721 caccccgaag caaagcccac ccttcgacgt ctccacccat gaattAacga ctgcgggcga
      781 ggacggaaga cgagtaaagt gtggtggaga gcggcgggga tcagcctagc tgccattccc
      841 agtgtacacg actgcaacga cgtcgtcgta ggctctgacc gtagtctccg aggtcctact
      901 tgatcagtac accgcccgca acaagctcga gctgttccca caccgcgctc acctgctgga
      961 ctgcctgcta ctgcttgctc agcgcacaca caccctcaaa gaacaccgtc gcaaaccgca
     1021 gcaccccgcc ggcgcgactg atggcatcac gaggaggacg aggaggaggg cgaggaggga
     1081 ggggaggagg ggcgtcgggg aagcctcagc cgtgactGAG ACCatgcATC TTTCTAGAAG
     1141 ATCTCCTACA ATATTCTCAG CTGCCATGGA AAATCGATGT TCTTCTTTTA TTCTCTCAAG
     1201 ATTTTCAGGC TGTATATTAA AACTTATATT AAGAACTATG CTAACCACCT CATCAGGAAC
     1261 CGTTGTAGGT GGCGTGGGTT TTCTTGGCAA TCGACTCTCA TGAAAACTAC GAGCTAAATA
     1321 TTCAATATGT TCCTCTTGAC CAACTTTATT CTGCATTTTT TTTGAACGAG GTTTAGAGCA
     1381 AGCTTCAGGA AACTGAGACA GGAATTTTAT TAAAAATTTA AATTTTGAAG AAAGTTCAGG
     1441 GTTAATAGCA TCCATTTTTT GCTTTGCAAG TTCCTCAGCA TTCTTAACAA AAGACGTCTC
     1501 TTTTGACATG TTTAAAGTTT AAACCTCCTG TGTGAAATTG TTATCCGCTC ACAATTCCAC
     1561 ACATTATACG AGCCGGAAGC ATAAAGTGTA AAGCCTGGGG TGCCTAATGA GTGAGCTAAC
     1621 TCACATTAAT TGCGTTGCGC TCACTGCCAA TTGCTTTCCA GTCGGGAAAC CTGTCGTGCC
     1681 AGCTGCATTA ATGAATCGGC CAACGCGCGG GGAGAGGCGG TTTGCGTATT GGGCGCTCTT
     1741 CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG
     1801 CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA
     1861 TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT
     1921 TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC
     1981 GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT
     2041 CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG
     2101 TGGCGCTTTC TCATAGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA
     2161 AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT
     2221 ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA
     2281 ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA
     2341 ACTACGGCTA CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT
     2401 TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT
     2461 TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA
     2521 TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA
     2581 TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT
     2641 CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG
     2701 CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT
     2761 AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG
     2821 ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC
     2881 GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG
     2941 CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA
     3001 TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA
     3061 GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA
     3121 TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA
     3181 ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA
     3241 AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG
     3301 ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG
     3361 GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG
     3421 CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG
     3481 GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC
     3541 TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA
     3601 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG
     3661 TGCCACCTGA CGTCTAAGAA ACCATTATTA TCATGACATT AACCTATAAA AATAGGCGTA
     3721 TCACGAGGCC
//