LOCUS       pJET1.2-tHe-Tub1A2      4488 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 2
FEATURES             Location/Qualifiers
     promoter        305..323
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: promoter for bacteriophage T7 RNA polymerase"
     Sequenced       353..1443
                     /label="1-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="Sequenced by SO501 (CGACTCACTATAGGGAGAGCGGC)"
     primer          372..393
                     /label="HeTub1C1-FWD"
                     /note="sequence: ggaaccttctctcctgtttctg"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    448..1797
                     /label="tHe-Tub1A2"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             448..1797
                     /label="Translation 448-1797"
                     /translation="MRECISIHVGQAGVQIGNACWELYCLEHGIQPDGQMPSDKTVGAADDSFNTFFSETGSGKHVPRAVFVDLEPTVVDEVRTGTYRQLFHPEQLITGKEDAANNYARGHYTVGKEIVDLVLDRVRKLADQCTGLQGFLIFHSFGGGTGSGFTSLLMERLSVDYGKKSKLEFSIYPAPQVSTAVVEPYNSILTTHTTLEHSDCAFMVNNEAIYDICRRNLDIERPTYTNLNRLIGQIVSSITASLRFDGALNVDLTEFQTNLVPYPRIHFPLVTYAPIISAEKAYHEQLTVSEITNSCFEPANQMVKCDPRHGKYMACCMLYRGDVVPKDVNAAIAAIKTKRSIQFVDWCPTGFKVGINYQPPTVVPGGDLAKVQRAVCMLSNTTAIAEAWARLDHKFDLMYAKRAFVHWYVGEGMEEGEFSEAREDLAALEKDYEEVGVDSVDAGEEGEEY*"
     sequencing      complement(1264..1890)
                     /label="1-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="confirmed by sequencing using SO511 (AAGAACATCGATTTTCCATGGCAG)"
     primer          complement(1864..1885)
                     /label="HeTub1C1-REV"
                     /note="sequence: gtttaaggacgagagattcccg"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    1886..1886
                     /label="Insertion site"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     protein_bind    2296..2312
                     /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(2320..2326)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2327..2344)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2345..2350)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2677..3265)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3436..4227)
                     /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(4228..4296)
                     /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(4297..4401)
                     /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 Tggaaccttc tctcctgttt ctgattcctc tacgtgtttt tttcgtctag
      421 tccacacaca cacagaccga atccaaaATG CGCGAATGCA TCTCTATCCA CGTCGGACAG
      481 GCCGGAGTGC AGATTGGCAA CGCCTGCTGG GAGCTTTACT GTCTGGAGCA TGGCATCCAG
      541 CCGGACGGGC AGATGCCGTC GGACAAGACG GTTGGCGCCG CTGACGACTC CTTCAACACC
      601 TTCTTCAGCG AGACCGGATC CGGCAAGCAC GTTCCGCGGG CCGTCTTCGT CGATCTCGAG
      661 CCGACCGTTG TTGATGAAGT GCGAACCGGA ACATACCGCC AGCTTTTCCA TCCAGAGCAG
      721 CTAATCACCG GAAAGGAGGA TGCCGCCAAC AATTACGCTC GCGGGCACTA CACCGTCGGC
      781 AAGGAGATTG TCGACCTGGT GCTGGACCGC GTGCGCAAGC TCGCCGACCA ATGTACCGGT
      841 CTGCAGGGCT TTCTCATCTT TCACTCGTTC GGCGGTGGGA CCGGATCCGG ATTCACCTCC
      901 CTGCTGATGG AACGCTTGTC GGTCGACTAC GGGAAGAAGT CCAAACTGGA ATTCTCCATC
      961 TACCCCGCCC CTCAGGTGTC GACTGCCGTG GTGGAGCCGT ACAACTCGAT CCTGACGACC
     1021 CATACCACGC TGGAGCACTC TGACTGTGCC TTCATGGTGA ACAACGAAGC CATCTACGAC
     1081 ATTTGCCGTC GGAACCTTGA CATTGAGCGT CCGACCTACA CCAACCTCAA CCGCCTGATC
     1141 GGGCAGATTG TCTCCTCCAT CACCGCGTCG CTGCGCTTCG ACGGCGCTCT CAACGTCGAT
     1201 CTCACCGAAT TCCAGACCAA CTTGGTTCCT TACCCCCGTA TCCATTTCCC GCTGGTGACC
     1261 TACGCCCCGA TCATTTCCGC TGAAAAGGCC TATCACGAAC AGTTGACGGT CTCGGAGATC
     1321 ACCAACTCGT GCTTCGAGCC GGCTAACCAG ATGGTCAAGT GCGACCCGCG CCACGGAAAG
     1381 TACATGGCCT GCTGCATGCT CTACCGCGGT GACGTTGTGC CCAAGGACGT GAACGCCGCC
     1441 ATCGCGGCCA TTAAGACCAA GCGCAGCATC CAGTTTGTCG ACTGGTGCCC GACTGGATTT
     1501 AAGGTGGGAA TCAACTATCA GCCCCCGACG GTGGTGCCGG GCGGGGATTT GGCCAAGGTG
     1561 CAGCGTGCCG TCTGCATGCT GTCCAACACC ACGGCCATCG CCGAAGCCTG GGCGCGTCTG
     1621 GACCACAAGT TCGACCTGAT GTACGCCAAG CGCGCCTTTG TCCACTGGTA CGTCGGTGAG
     1681 GGCATGGAGG AGGGCGAGTT CAGCGAGGCC CGTGAAGATT TGGCCGCCCT CGAGAAGGAC
     1741 TACGAGGAGG TCGGTGTGGA CTCTGTCGAC GCTGGTGAGG AAGGCGAGGA ATATTGAgac
     1801 cactcctctc gtttcccttc tttaatctct ggcccttttt tctctcgctc gatacatttg
     1861 actcgggaat ctctcgtcct taaacATCTT TCTAGAAGAT CTCCTACAAT ATTCTCAGCT
     1921 GCCATGGAAA ATCGATGTTC TTCTTTTATT CTCTCAAGAT TTTCAGGCTG TATATTAAAA
     1981 CTTATATTAA GAACTATGCT AACCACCTCA TCAGGAACCG TTGTAGGTGG CGTGGGTTTT
     2041 CTTGGCAATC GACTCTCATG AAAACTACGA GCTAAATATT CAATATGTTC CTCTTGACCA
     2101 ACTTTATTCT GCATTTTTTT TGAACGAGGT TTAGAGCAAG CTTCAGGAAA CTGAGACAGG
     2161 AATTTTATTA AAAATTTAAA TTTTGAAGAA AGTTCAGGGT TAATAGCATC CATTTTTTGC
     2221 TTTGCAAGTT CCTCAGCATT CTTAACAAAA GACGTCTCTT TTGACATGTT TAAAGTTTAA
     2281 ACCTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC ATTATACGAG CCGGAAGCAT
     2341 AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG CGTTGCGCTC
     2401 ACTGCCAATT GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA
     2461 ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC TCACTGACTC
     2521 GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG
     2581 GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG GCCAGCAAAA
     2641 GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA
     2701 CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG
     2761 ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT
     2821 TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG
     2881 CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC
     2941 CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT
     3001 AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA
     3061 TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGGAC
     3121 AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC
     3181 TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT
     3241 TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG GGTCTGACGC
     3301 TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT
     3361 CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA
     3421 AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT
     3481 ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA TACGGGAGGG
     3541 CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC CCACGCTCAC CGGCTCCAGA
     3601 TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC CTGCAACTTT
     3661 ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT
     3721 TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC GCTCGTCGTT
     3781 TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT GATCCCCCAT
     3841 GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA GTAAGTTGGC
     3901 CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG TCATGCCATC
     3961 CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG AATAGTGTAT
     4021 GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT AATACCGCGC CACATAGCAG
     4081 AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT CAAGGATCTT
     4141 ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT CTTCAGCATC
     4201 TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG CCGCAAAAAA
     4261 GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC AATATTATTG
     4321 AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA TTTAGAAAAA
     4381 TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG TCTAAGAAAC
     4441 CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC ACGAGGCC
//