LOCUS       pJET1.2-tHe-Tub2D3      4335 bp ds-DNA     circular     26-APR-2023
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. colony 11b
FEATURES             Location/Qualifiers
     promoter        305..323
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     misc_feature    346..1424
                     /label="11b-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          372..392
                     /label="HeTub2D3-FWD"
                     /note="sequence: gcaagcATGCGTGAAATCGTC"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             378..1721
                     /label="tHe-Tub2D3"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             378..1721
                     /label="Translation 378-1721"
                     /translation="MREIVHLQAGQCGNQIGAKFWEVISDEHGIDPTGTYHGDSDLQLERINVYYNEATGGKYVPRAVLVDLEPGTMDSVRSGPFGQLFRPDNFIFGLSGAGNNWAKGHYTEGAELVDSVLDVVRKEAESCDCLQGFQLTHSLGGGTGSGMGTLMISKIREEYPDRIMMTFSVVPSPKVSDTVVEPYNATLSVHQLVENTDETFCIDNEALYDICFRTLKLTTPTYGDLNHLVSATMSGVTTCLRFPGQLNADLRKLAVNMVPFPRLHFFMPGFAPLTSRGSQQYRALTVPELTQQMFDAKNMMAACDPRHGRYLTVAAIFRGRMSMKEVDEQLLNIQNKNSSYFVEWIPNNVKTAVCDIPPRGLKMSATFIGNSTAIQELFKRVSEQFTAMFRRKAFLHWYTGEGMDEMEFTEAESNMNDLVSEYQQYQEATAEDEGEFEEEGEGGEAEA*"
     misc_feature    complement(734..1737)
                     /label="11b-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(1711..1732)
                     /label="HeTub2D3-REV"
                     /note="sequence: ggtccttccaaCTAAGCCTCAG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1733..1733
                     /label="Insertion site"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     protein_bind    2143..2159
                     /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(2167..2173)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2174..2191)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2192..2197)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2524..3112)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3283..4074)
                     /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(4075..4143)
                     /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(4144..4248)
                     /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 TgcaagcATG CGTGAAATCG TCCATTTGCA AGCGGGCCAA TGTGGAAACC
      421 AAATCGGAGC GAAGTTTTGG GAAGTCATCT CGGACGAGCA TGGCATCGAT CCCACCGGCA
      481 CGTACCACGG AGACTCCGAT CTCCAGTTGG AGAGGATCAA CGTTTATTAC AATGAAGCCA
      541 CCGGAGGGAA ATATGTGCCG CGCGCGGTTC TGGTGGATCT CGAGCCGGGA ACGATGGACT
      601 CTGTGCGGTC CGGGCCATTT GGACAGCTTT TCCGTCCGGA CAACTTTATT TTCGGCCTGA
      661 GCGGAGCCGG AAACAACTGG GCCAAGGGTC ACTACACCGA GGGCGCCGAA CTGGTGGACT
      721 CGGTCCTCGA CGTGGTCCGC AAGGAGGCCG AGAGCTGCGA CTGTCTGCAG GGCTTCCAGC
      781 TGACTCACTC TCTCGGCGGT GGAACCGGAT CGGGAATGGG CACCCTGATG ATCTCCAAGA
      841 TCCGTGAGGA GTATCCCGAT CGCATCATGA TGACCTTCAG TGTCGTCCCA TCTCCCAAGG
      901 TGTCCGACAC TGTGGTGGAG CCGTACAACG CCACCCTGTC CGTCCACCAG CTGGTGGAGA
      961 ACACGGACGA GACCTTCTGC ATCGACAATG AGGCGCTCTA CGACATCTGC TTCCGCACTC
     1021 TCAAGCTGAC CACACCAACG TACGGCGACT TGAACCATCT CGTCTCGGCG ACCATGTCCG
     1081 GTGTGACTAC CTGCCTGCGC TTCCCGGGTC AGCTGAACGC CGATCTCCGC AAACTGGCCG
     1141 TCAACATGGT CCCCTTCCCG CGGCTGCACT TCTTCATGCC CGGCTTCGCC CCGCTGACCT
     1201 CGCGTGGTTC CCAGCAGTAC CGCGCGCTGA CCGTGCCGGA ACTGACCCAG CAGATGTTCG
     1261 ACGCGAAGAA TATGATGGCC GCCTGCGATC CCCGCCACGG ACGCTACCTC ACCGTGGCGG
     1321 CAATCTTCCG CGGCCGGATG AGCATGAAGG AGGTGGATGA ACAGCTACTG AACATCCAGA
     1381 ACAAGAACAG CAGCTACTTC GTCGAGTGGA TCCCCAACAA CGTGAAGACC GCGGTCTGCG
     1441 ACATTCCACC GCGCGGTTTG AAGATGTCCG CCACCTTCAT CGGAAACAGC ACTGCCATCC
     1501 AGGAGCTGTT CAAACGCGTT TCGGAACAGT TCACTGCCAT GTTCCGCCGA AAAGCGTTCC
     1561 TGCATTGGTA CACTGGAGAG GGTATGGACG AGATGGAATT CACCGAAGCC GAGTCCAACA
     1621 TGAATGACTT GGTGTCGGAG TACCAACAGT ACCAGGAGGC AACCGCTGAA GACGAGGGCG
     1681 AATTCGAGGA GGAAGGCGAG GGTGGAGAAG CTGAGGCTTA Gttggaagga ccATCTTTCT
     1741 AGAAGATCTC CTACAATATT CTCAGCTGCC ATGGAAAATC GATGTTCTTC TTTTATTCTC
     1801 TCAAGATTTT CAGGCTGTAT ATTAAAACTT ATATTAAGAA CTATGCTAAC CACCTCATCA
     1861 GGAACCGTTG TAGGTGGCGT GGGTTTTCTT GGCAATCGAC TCTCATGAAA ACTACGAGCT
     1921 AAATATTCAA TATGTTCCTC TTGACCAACT TTATTCTGCA TTTTTTTTGA ACGAGGTTTA
     1981 GAGCAAGCTT CAGGAAACTG AGACAGGAAT TTTATTAAAA ATTTAAATTT TGAAGAAAGT
     2041 TCAGGGTTAA TAGCATCCAT TTTTTGCTTT GCAAGTTCCT CAGCATTCTT AACAAAAGAC
     2101 GTCTCTTTTG ACATGTTTAA AGTTTAAACC TCCTGTGTGA AATTGTTATC CGCTCACAAT
     2161 TCCACACATT ATACGAGCCG GAAGCATAAA GTGTAAAGCC TGGGGTGCCT AATGAGTGAG
     2221 CTAACTCACA TTAATTGCGT TGCGCTCACT GCCAATTGCT TTCCAGTCGG GAAACCTGTC
     2281 GTGCCAGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGGCG
     2341 CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT
     2401 ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA
     2461 GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC
     2521 GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG
     2581 GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT
     2641 GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG
     2701 AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG
     2761 CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG
     2821 TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC
     2881 TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG
     2941 GCCTAACTAC GGCTACACTA GAAGGACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT
     3001 TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG
     3061 TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC
     3121 TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT
     3181 GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT
     3241 TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG
     3301 TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT
     3361 CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC
     3421 GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC
     3481 CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG
     3541 GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC
     3601 AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG
     3661 ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC
     3721 TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT
     3781 GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC
     3841 AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT
     3901 ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC
     3961 TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC
     4021 TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA
     4081 AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT
     4141 CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG
     4201 ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG
     4261 AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG ACATTAACCT ATAAAAATAG
     4321 GCGTATCACG AGGCC
//