LOCUS       pJET1.2-tHe-Tub2D4      4363 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. clone 2C
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    352..1439
                     /label="2c-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             398..1744
                     /label="tHe-Tub2D4"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             398..1744
                     /label="Translation 398-1744"
                     /translation="MREIVHLQVGQCGNQIGAKFWEVISDEHGIDPTGTYRGDSDLQLERINVYYNEAGNNKYVPRAVLVDLEPGTMDSVRSSPFGQLFRPDNFIFGQSGAGNNWAKGHYTEGAELVDSVLDVVRKEAENCDCLQGFQLTHSLGGGTGSGMGTLLISKIREEYPDRIMTTFSVVPSPKVSDTVVEPYNATLSVHQLVENTDETFCIDNEALYDICFRTLKLTTPTYGDLNHLVSATMSGVTTCFRFPGQLNADLRKLAVNMVPFPRLHFFMPGFAPLTARGSQGYRALSVAELTTQMFDSKNMMAACDPRHGRYLTVAAIFRGRMSMKEVDDQMLQIQNKNSSYFVEWIPNNIKTAVCDIPPRGLKMAATFIGNSTAIQELFKRISEQFTAMFRRKAFLHWYTGEGMDEMEFTEAESNMNDLISEYQQYQEATAEDEGEFEEGEQEAQEEAA*"
     misc_feature    complement(786..1765)
                     /label="2c-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="verified by sequencing"
     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"
     CDS             complement(3311..4102)
                     /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(4103..4171)
                     /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(4172..4276)
                     /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 Tcgcttcaga aacccttatc agcaaagATG CGCGAAATCG TCCACCTCCA
      421 AGTCGGCCAG TGCGGCAACC AGATCGGAGC GAAGTTCTGG GAAGTGATCT CCGACGAACA
      481 TGGCATCGAC CCCACCGGCA CCTACCGCGG CGACTCGGAT TTGCAGTTGG AACGGATCAA
      541 TGTTTACTAT AACGAAGCTG GAAACAACAA ATATGTTCCG CGCGCTGTTT TGGTCGATCT
      601 TGAGCCGGGA ACGATGGATT CGGTCCGATC ATCTCCGTTT GGTCAaCTTT TCCGACCGGA
      661 TAACTTTATc TTTGGCCAGA GTGGTGCGGG AAACAACTGG GCTAAAGGCC ACTACACCGA
      721 GGGTGCCGAA CTGGTGGACT CTGTTCTCGA TGTCGTGCGC AAGGAGGCCG AGAACTGCGA
      781 CTGCCTCCAG GGATTTCAGC TGACTCACTC GCTGGGCGGC GGCACTGGAT CCGGTATGGG
      841 tACTCTGCTC ATCTCtAAGA TCCGCGAGGA GTACCCCGAT CGCATCATGA CCACATTCAG
      901 CGTTGTTCCA TCGCCGAAGG TGTCTGACAC TGTGGTGGAG CCCTACAACG CGACTCTGTC
      961 GGTCCACCAG CTGGTCGAGA ATACCGACGA GACGTTTTGC ATCGACAACG AAGCGCTGTA
     1021 CGACATCTGC TTCCGCACCC TCAAGCTGAC CACCCCGACG TACGGCGACC TGAACCACCT
     1081 CGTGTCTGCC ACCATGTCGG GTGTGACGAC CTGCTTCCGT TTCCCGGGCC AACTGAACGC
     1141 CGATCTCCGC AAGTTGGCCG TCAACATGGT GCCCTTCCCG CGGCTGCACT TTTTCATGCC
     1201 CGGTTTCGCC CCGTTGACAG CTCGCGGCAG CCAGGGCTAC CGTGCCCTCA GCGTCGCCGA
     1261 ACTCACCACC CAGATGTTCG ACTCGAAGAA CATGATGGCC GCCTGCGATC CCCGTCACGG
     1321 ACGCTACCTC ACCGTGGCGG CCATCTTCCG CGGCCGGATG AGCATGAAGG AAGTGGACGA
     1381 CCAAATGCTG CAGATCCAGA ACAAGAACTC CAGCTACTTC GTTGAATGGA TTCCCAACAA
     1441 CATCAAGACA GCCGTCTGCG ACATCCCTCC GCGCGGTCTG AAGATGGCGG CCACGTTCAT
     1501 CGGCAACAGC ACTGCCATCC AGGAGCTGTT CAAGCGCATC AGCGAGCAGT TCACTGCCAT
     1561 GTTCCGTCGC AAGGCCTTCC TCCATTGGTA CACCGGCGAG GGCATGGACG AGATGGAATT
     1621 CACCGAGGCC GAGAGCAACA TGAACGATCT CATCTCGGAG TACCAGCAGT ACCAGGAGGC
     1681 CACCGCcGAG GATGAGGGTG AATTTGAGGA GGGCGAGCAG GAGGCCCAGG AGGAgGCTGC
     1741 TTAGactagt ttctcgctct 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 AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT
     3301 GGTCTGACAG TTACCAATGC TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC
     3361 GTTCATCCAT AGTTGCCTGA CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC
     3421 CATCTGGCCC CAGTGCTGCA ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT
     3481 CAGCAATAAA CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG
     3541 CCTCCATCCA GTCTATTAAT TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA
     3601 GTTTGCGCAA CGTTGTTGCC ATTGCTACAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA
     3661 TGGCTTCATT CAGCTCCGGT TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT
     3721 GCAAAAAAGC GGTTAGCTCC TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG
     3781 TGTTATCACT CATGGTTATG GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA
     3841 GATGCTTTTC TGTGACTGGT GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC
     3901 GACCGAGTTG CTCTTGCCCG GCGTCAATAC GGGATAATAC CGCGCCACAT AGCAGAACTT
     3961 TAAAAGTGCT CATCATTGGA AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC
     4021 TGTTGAGATC CAGTTCGATG TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA
     4081 CTTTCACCAG CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA
     4141 TAAGGGCGAC ACGGAAATGT TGAATACTCA TACTCTTCCT TTTTCAATAT TATTGAAGCA
     4201 TTTATCAGGG TTATTGTCTC ATGAGCGGAT ACATATTTGA ATGTATTTAG AAAAATAAAC
     4261 AAATAGGGGT TCCGCGCACA TTTCCCCGAA AAGTGCCACC TGACGTCTAA GAAACCATTA
     4321 TTATCATGAC ATTAACCTAT AAAAATAGGC GTATCACGAG GCC
//