LOCUS       pJET1.2-tHe-Tub1C1      4526 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 6
FEATURES             Location/Qualifiers
     promoter        305..323
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     Sequencing      351..1389
                     /label="3-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          372..393
                     /label="HeTub1C2-FWD"
                     /note="sequence: cccatacagctcacaagattcc"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     CDS             444..1802
                     /label="tHe-Tub1C1"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             444..1802
                     /label="Translation 444-1802"
                     /translation="MRECISIHVGQAGVQIGNACWELYCLEHGIQPDGRMSQSKEGGKRDGQDSFTTFFSETGSGKYVPRAVFVDLEPTVVDEVRTGTYKQLFHPEQLITGKEDAANNYARGHYTVGKEIVDVTIDRIRKLADQCTGLQGFLIFHSFGGGTGSGFTSLLMERLSVEFGKKSKLEFCVYPAPEVSTAVVEPYNSILTTHTTLEHSDCAFMVDNEAIFDICRRKLDVERPTYTNLNRLISQIVSSITASLRFDGALNVDLTEFQTNLVPYPRIHFPLVSYSPVISAEKAFHEQLSVQEITNACFEPANQMVKCDPRQGKYMACCMLYRGDVVPKDVTAAIATVKSKSSIRFVDWCPTGFKVGLNFQPPTVVPGADQAAVPRAVCMLSNTTAIVEAWARLNHKFDLMYAKRAFVHWYVGEGMEEGEFSEAREDLAALERDYEEAGADSTDGQDDENGEF*"
     Sequencing      complement(1024..1928)
                     /label="3-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(1898..1923)
                     /label="HeTub1C2-REV"
                     /note="sequence: ccactcgcttcccatgataaaacaag"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1924..1924
                     /label="Insertion site"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     protein_bind    2334..2350
                     /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(2358..2364)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2365..2382)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2383..2388)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2715..3303)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3474..4265)
                     /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(4266..4334)
                     /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(4335..4439)
                     /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 Tcccatacag ctcacaagat tcccttctta atatcaaact atttgcggat
      421 ctatttcggc gacctcaaAg aaaATGCGTG AATGTATCTC AATCCACGTC GGACAGGCTG
      481 GAGTGCAGAT CGGCAATGCC TGCTGGGAAC TGTACTGCCT GGAGCACGGC ATCCAGCCGG
      541 ACGGACGGAT GTCGCAGTCG AAGGAAGGCG GCAAACGTGA CGGCCAAGAC TCATTCACCA
      601 CTTTCTTCAG CGAAACTGGG TCAGGCAAAT ACGTTCCCCG AGCTGTTTTT GTGGACCTTG
      661 AGCCTACTGT TGTCGACGAA GTTCGGACAG GCACCTACAA GCAGCTTTTC CATCCGGAGC
      721 AGCTGATTAC CGGGAAGGAG GATGCCGCCA ACAACTACGC TCGTGGCCAT TACACCGTCG
      781 GCAAGGAGAT TGTCGACGTC ACCATTGACC GGATCCGGAA GTTGGCTGAC CAGTGCACTG
      841 GCCTGCAGGG ATTTCTTATT TTTCATTCGT TCGGCGGTGG CACTGGTTCG GGTTTCACTT
      901 CGTTGCTCAT GGAACGCTTG TCTGTCGAGT TTGGCAAGAA GTCAAAGTTG GAGTTTTGCG
      961 TCTACCCCGC TCCGGAAGTC TCGACTGCTG TGGTTGAACC GTACAATTCC ATCCTCACCA
     1021 CCCACACCAC CCTGGAGCAC TCGGACTGTG CATTCATGGT AGACAATGAA GCCATTTTTG
     1081 ACATCTGTCG CCGGAAATTG GATGTCGAGC GACCGACTTA CACCAACTTG AACCGGCTCA
     1141 TCAGCCAGAT CGTTTCTTCC ATCACAGCCT CGCTCCGCTT CGACGGCGCC CTGAACGTCG
     1201 ACCTCACGGA ATTCCAGACC AACTTGGTGC CGTATCCGCG TATCCACTTC CCGCTGGTAT
     1261 CTTACTCACC CGTCATTTCA GCGGAAAAGG CGTTCCACGA GCAGTTGTCG GTCCAAGAGA
     1321 TCACCAATGC CTGCTTCGAG CCGGCCAACC AGATGGTGAA GTGCGACCCG CGCCAGGGCA
     1381 AATACATGGC CTGCTGCATG CTCTACCGCG GTGATGTGGT TCCGAAGGAT GTCACCGCGG
     1441 CCATCGCGAC CGTGAAGAGC AAGTCGAGCA TCCGTTTCGT GGATTGGTGC CCGACCGGTT
     1501 TCAAGGTTGG CTTGAACTTC CAGCCACCGA CCGTCGTCCC GGGCGCCGAT CAGGCAGCCG
     1561 TTCCGCGTGC CGTTTGCATG CTTTCCAACA CCACCGCCAT CGTGGAAGCC TGGGCGAGGT
     1621 TGAACCACAA GTTCGACCTG ATGTACGCCA AGCGCGCCTT TGTCCACTGG TACGTCGGCG
     1681 AGGGCATGGA AGAGGGCGAG TTCAGCGAGG CCCGAGAAGA TTTGGCCGCC CTCGAGAGGG
     1741 ACTACGAGGA GGCCGGCGCG GACTCTACCG ACGGCCAGGA CGACGAAAAT GGCGAATTCT
     1801 AAagccggta tcaccagagc tgtatttttg tcgaactttc ggattttcgt cgctatgttt
     1861 tcgtttttat cctgtcctat gttttctaat taatatactt gttttatcat gggaagcgag
     1921 tggATCTTTC TAGAAGATCT CCTACAATAT TCTCAGCTGC CATGGAAAAT CGATGTTCTT
     1981 CTTTTATTCT CTCAAGATTT TCAGGCTGTA TATTAAAACT TATATTAAGA ACTATGCTAA
     2041 CCACCTCATC AGGAACCGTT GTAGGTGGCG TGGGTTTTCT TGGCAATCGA CTCTCATGAA
     2101 AACTACGAGC TAAATATTCA ATATGTTCCT CTTGACCAAC TTTATTCTGC ATTTTTTTTG
     2161 AACGAGGTTT AGAGCAAGCT TCAGGAAACT GAGACAGGAA TTTTATTAAA AATTTAAATT
     2221 TTGAAGAAAG TTCAGGGTTA ATAGCATCCA TTTTTTGCTT TGCAAGTTCC TCAGCATTCT
     2281 TAACAAAAGA CGTCTCTTTT GACATGTTTA AAGTTTAAAC CTCCTGTGTG AAATTGTTAT
     2341 CCGCTCACAA TTCCACACAT TATACGAGCC GGAAGCATAA AGTGTAAAGC CTGGGGTGCC
     2401 TAATGAGTGA GCTAACTCAC ATTAATTGCG TTGCGCTCAC TGCCAATTGC TTTCCAGTCG
     2461 GGAAACCTGT CGTGCCAGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG
     2521 CGTATTGGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG
     2581 CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT
     2641 AACGCAGGAA AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC
     2701 GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC
     2761 TCAAGTCAGA GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA
     2821 AGCTCCCTCG TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT
     2881 CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG
     2941 TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC
     3001 GCCTTATCCG GTAACTATCG TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG
     3061 GCAGCAGCCA CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC
     3121 TTGAAGTGGT GGCCTAACTA CGGCTACACT AGAAGGACAG TATTTGGTAT CTGCGCTCTG
     3181 CTGAAGCCAG TTACCTTCGG AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC
     3241 GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT
     3301 CAAGAAGATC CTTTGATCTT TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT
     3361 TAAGGGATTT TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA
     3421 AAATGAAGTT TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA
     3481 TGCTTAATCA GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC
     3541 TGACTCCCCG TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT
     3601 GCAATGATAC CGCGAGACCC ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA
     3661 GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT
     3721 AATTGTTGCC GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT
     3781 GCCATTGCTA CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC
     3841 GGTTCCCAAC GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC
     3901 TCCTTCGGTC CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT
     3961 ATGGCAGCAC TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT
     4021 GGTGAGTACT CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC
     4081 CCGGCGTCAA TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT
     4141 GGAAAACGTT CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG
     4201 ATGTAACCCA CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT
     4261 GGGTGAGCAA AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA
     4321 TGTTGAATAC TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT
     4381 CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC
     4441 ACATTTCCCC GAAAAGTGCC ACCTGACGTC TAAGAAACCA TTATTATCAT GACATTAACC
     4501 TATAAAAATA GGCGTATCAC GAGGCC
//