LOCUS       pJET1.2-tHe-Tub3        4393 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 16b
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    351..1367
                     /label="16b-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          372..395
                     /label="HeTub3-FWD"
                     /note="sequence: ATGCCTCGCTCCATCATCTCCCTG"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             372..1754
                     /label="tHe-Tub3"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             372..1754
                     /label="Translation 372-1754"
                     /translation="MPRSIISLQLGQCGNQVGMEFWKKLCMEHGIESNGQLKNSGAEGDFRDRKDFFFYQADDSHYIPRAILIDLEPRVIQTIKNSDYGKLFNPENIYVSEQGGGAGNNWAAGFGMGQDVSEKVFDMIEREAEGCDGLEGFILSHSIAGGTGSGMGSYVLEHLHDRFPKKLIQTFSIFPNQDDSASDVVVQPYNSILTMRRLAENADCVVVLDNTALNSIAAERLRIDQPSLTEINNMVSTVMAASTSTLRYPGYMYNNFVGLMAPLIPMPKLHFLMAGYAPLTSLDQTSGGIQKTTVFDVMRRLLQPKNMMLSPGKDALAADNRYLSILNIIQGEVESGEIHKSLMRIKERNLVKFMEWGPASIHVAQSKRSPFVASNHRISGLMLANHTSVQGLFDSIVKQYDKMKKRNAYLDRFTNEKNWAGVSDMEQSREVVANLIEEYKAAAKPDYIDYLRGKLQSKKM*"
     misc_feature    complement(894..1795)
                     /label="16b-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(1769..1791)
                     /label="HeTub3-REV1"
                     /note="sequence: tgggacgatgtaaagggtgaagg"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     protein_bind    2201..2217
                     /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(2225..2231)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2232..2249)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2250..2255)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2582..3170)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3341..4132)
                     /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(4133..4201)
                     /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(4202..4306)
                     /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 TATGCCTCGC TCCATCATCT CCCTGCAGCT CGGCCAGTGC GGCAACCAGG
      421 TCGGCATGGA ATTCTGGAAG AAATTGTGCA TGGAGCATGG CATCGAGAGC AACGGCCAGC
      481 TGAAGAATTC CGGCGCGGAG GGCGACTTTC GGGATAGGAA GGATTTCTTC TTCTACCAGG
      541 CCGACGACAG CCACTACATT CCGCGCGCCA TCCTCATCGA TCTCGAGCCG CGTGTGATTC
      601 AGACCATCAA GAATTCCGAC TACGGCAAGC TGTTTAATCC GGAAAACATT TATGTGTCGG
      661 AGCAGGGCGG TGGTGCGGGA AACAACTGGG CTGCAGGATT CGGAATGGGG CAAGACGTCT
      721 CGGAGAAAGT GTTCGACATG ATTGAGCGTG AAGCGGAAGG CTGCGACGGA TTAGAGGGCT
      781 TCATTCTCAG TCACTCCATT GCCGGCGGTA CCGGGTCCGG CATGGGCTCA TACGTCCTGG
      841 AGCATTTGCA TGATCGTTTC CCGAAGAAGC TCATTCAGAC CTTCTCCATC TTCCCCAATC
      901 AGGACGACTC TGCCAGCGAT GTGGTGGTTC AGCCGTACAA CTCGATCTTG ACGATGCGCC
      961 GGCTGGCGGA GAACGCCGAC TGTGTGGTGG TGCTGGACAA CACGGCGCTC AATTCGATCG
     1021 CTGCAGAACG GCTGCGAATC GACCAACCGA GCTTGACAGA GATCAACAAT ATGGTCTCAA
     1081 CGGTGATGGC CGCCAGTACG TCCACATTGC GCTATCCCGG TTACATGTAC AACAACTTTG
     1141 TCGGCTTGAT GGCCCCGCTC ATTCCCATGC CGAAGCTCCA TTTTCTCATG GCCGGCTATG
     1201 CTCCACTGAC GTCGTTGGAC CAGACGAGCG GTGGCATCCA GAAAACGACG GTATTTGACG
     1261 TTATGCGTCG TTTACTTCAG CCGAAGAACA TGATGCTGTC CCCGGGGAAG GATGCCCTGG
     1321 CTGCCGACAA TCGCTATCTC TCCATTTTGA ACATCATTCA GGGCGAAGTG GAATCTGGCG
     1381 AGATTCACAA AAGCCTGATG CGCATTAAAG AACGCAATCT GGTCAAGTTT ATGGAATGGG
     1441 GTCCGGCCAG CATCCATGTG GCCCAGTCGA AGCGCTCGCC GTTTGTCGCG TCCAACCACC
     1501 GCATCAGCGG ACTGATGTTG GCCAACCACA CCAGCGTGCA GGGACTCTTC GACAGCATCG
     1561 TCAAGCAGTA CGACAAGATG AAGAAGCGCA ATGCCTACTT GGATCGCTTC ACCAATGAGA
     1621 AGAATTGGGC CGGTGTGTCG GACATGGAGC AGTCGAGGGA GGTGGTGGCC AACCTGATTG
     1681 AGGAGTACAA GGCAGCCGCG AAACCGGATT ACATAGACTA TCTGCGGGGG AAACTGCAGA
     1741 gtaagaagat gtgaagggat ccttggctcc ttcacccttt acatcgtccc aTCTTTCTAG
     1801 AAGATCTCCT ACAATATTCT CAGCTGCCAT GGAAAATCGA TGTTCTTCTT TTATTCTCTC
     1861 AAGATTTTCA GGCTGTATAT TAAAACTTAT ATTAAGAACT ATGCTAACCA CCTCATCAGG
     1921 AACCGTTGTA GGTGGCGTGG GTTTTCTTGG CAATCGACTC TCATGAAAAC TACGAGCTAA
     1981 ATATTCAATA TGTTCCTCTT GACCAACTTT ATTCTGCATT TTTTTTGAAC GAGGTTTAGA
     2041 GCAAGCTTCA GGAAACTGAG ACAGGAATTT TATTAAAAAT TTAAATTTTG AAGAAAGTTC
     2101 AGGGTTAATA GCATCCATTT TTTGCTTTGC AAGTTCCTCA GCATTCTTAA CAAAAGACGT
     2161 CTCTTTTGAC ATGTTTAAAG TTTAAACCTC CTGTGTGAAA TTGTTATCCG CTCACAATTC
     2221 CACACATTAT ACGAGCCGGA AGCATAAAGT GTAAAGCCTG GGGTGCCTAA TGAGTGAGCT
     2281 AACTCACATT AATTGCGTTG CGCTCACTGC CAATTGCTTT CCAGTCGGGA AACCTGTCGT
     2341 GCCAGCTGCA TTAATGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT ATTGGGCGCT
     2401 CTTCCGCTTC CTCGCTCACT GACTCGCTGC GCTCGGTCGT TCGGCTGCGG CGAGCGGTAT
     2461 CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC AGGGGATAAC GCAGGAAAGA
     2521 ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT
     2581 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT
     2641 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC
     2701 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA
     2761 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT
     2821 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA
     2881 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG
     2941 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC
     3001 CTAACTACGG CTACACTAGA AGGACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA
     3061 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG
     3121 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT
     3181 TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA CTCACGTTAA GGGATTTTGG
     3241 TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT AAATTAAAAA TGAAGTTTTA
     3301 AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG TTACCAATGC TTAATCAGTG
     3361 AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT AGTTGCCTGA CTCCCCGTCG
     3421 TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC CAGTGCTGCA ATGATACCGC
     3481 GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA CCAGCCAGCC GGAAGGGCCG
     3541 AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA GTCTATTAAT TGTTGCCGGG
     3601 AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA CGTTGTTGCC ATTGCTACAG
     3661 GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT CAGCTCCGGT TCCCAACGAT
     3721 CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC GGTTAGCTCC TTCGGTCCTC
     3781 CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT CATGGTTATG GCAGCACTGC
     3841 ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC TGTGACTGGT GAGTACTCAA
     3901 CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG CTCTTGCCCG GCGTCAATAC
     3961 GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT CATCATTGGA AAACGTTCTT
     4021 CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC CAGTTCGATG TAACCCACTC
     4081 GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG CGTTTCTGGG TGAGCAAAAA
     4141 CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC ACGGAAATGT TGAATACTCA
     4201 TACTCTTCCT TTTTCAATAT TATTGAAGCA TTTATCAGGG TTATTGTCTC ATGAGCGGAT
     4261 ACATATTTGA ATGTATTTAG AAAAATAAAC AAATAGGGGT TCCGCGCACA TTTCCCCGAA
     4321 AAGTGCCACC TGACGTCTAA GAAACCATTA TTATCATGAC ATTAACCTAT AAAAATAGGC
     4381 GTATCACGAG GCC
//