LOCUS       pJET1.2-tHe-Tub5        4379 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 19b
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..1420
                     /label="19b-F-seq"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          372..403
                     /label="HeTub5-FWD"
                     /note="sequence: gcaattcaaATGTCGCATAATGTGGTCATCTC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             381..1772
                     /label="tHe-Tub5"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             381..1772
                     /label="Translation 381-1772"
                     /translation="MSHNVVISIGQCGNQIGSEFWDRVVHSHALLDTDGTFTSALATFFRNTDGRHDSSLPIGSRLKHLQANAVLIDMETRVVDRLLRGPLGDLFDKRMTLTDSCGSGNNWSTGHLTYGLQHEDTISEMLRRSAEPLDQLQSFSVLHSMGGGTGSGLGTAVVKMMARDYGKTARLVSCVFPGDQDDVVTSPYNSVLALKVLAEDADSVISLDNNALSTVCSKHVLAGRPLRKSPPVPFDQMNDLVARMLADLTSSIRFGGELSIPLSELPVRMSPFPRMKFLIPGLSPVTGALPNMESKKIDRTFYEAFERRNQLMDCDPQRSAYLACLLLARGKDIAPSDVQRNVDKMKRYIHFAPGSKDEWAVSLCGIPSQSHPRSFLTLANNGCFHKPMQCLKERFVAMYRGRAHLHHYLLNGMEAADFEASLESLSALISDYRQVDAAGAVAANVRDRGYVAVGVVKPKVKAG*"
     misc_feature    complement(820..1781)
                     /label="19b-R-seq"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(1754..1776)
                     /label="HeTub5-REV"
                     /note="sequence: actgTCAGCCCGCCTTCACCTTG"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    1777..1777
                     /label="Insertion site"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     protein_bind    2187..2203
                     /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(2211..2217)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2218..2235)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     promoter        complement(2236..2241)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(2568..3156)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="direction: LEFT note: high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(3327..4118)
                     /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(4119..4187)
                     /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(4188..4292)
                     /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 Tgcaattcaa ATGTCGCATA ATGTGGTCAT CTCGATCGGG CAGTGCGGCA
      421 ATCAAATTGG ATCAGAGTTT TGGGATCGCG TCGTCCATTC ACACGCCTTA CTGGACACCG
      481 ATGGGACTTT CACTTCAGCG CTTGCCACTT TTTTCCGCAA CACCGATGGC CGACACGATT
      541 CGAGTCTACC TATCGGTAGT CGACTCAAGC ATTTGCAGGC CAACGCCGTG CTTATCGACA
      601 TGGAAACTCG CGTGGTGGAC AGGCTCCTCC GCGGCCCGTT GGGTGATCTG TTTGACAAGC
      661 GGATGACGTT GACAGACTCC TGTGGAAGCG GGAACAACTG GTCCACTGGA CATCTGACCT
      721 ACGGCCTGCA ACATGAAGAC ACGATTTCGG AGATGCTCCG GAGAAGTGCA GAGCCGCTTG
      781 ACCAGCTGCA GTCTTTTTCA GTCCTTCACT CTATGGGCGG TGGAACTGGC AGTGGCTTGG
      841 GTACGGCCGT GGTGAAGATG ATGGCACGGG ATTATGGCAA AACGGCGCGG TTGGTCAGTT
      901 GCGTTTTTCC CGGGGATCAG GATGATGTCG TCACGTCGCC GTACAATTCT GTTCTGGCTC
      961 TGAAGGTATT GGCTGAAGAC GCTGATAGCG TGATTAGCCT CGACAACAAC GCCCTCTCCA
     1021 CAGTCTGTTC AAAACACGTT CTCGCCGGCC GGCCACTTCG AAAAAGTCCT CCCGTTCCGT
     1081 TCGATCAAAT GAACGACTTG GTAGCCCGGA TGTTGGCCGA CCTGACCAGT TCCATTCGCT
     1141 TCGGCGGCGA ACTCAGTATT CCGCTGAGCG AGCTCCCCGT CCGGATGAGC CCGTTCCCGC
     1201 GGATGAAATT CCTCATACCC GGATTGAGTC CCGTGACTGG AGCGCTGCCC AATATGGAGT
     1261 CGAAGAAGAT TGACCGGACC TTCTACGAAG CGTTCGAGCG ACGCAATCAA CTGATGGACT
     1321 GCGACCCGCA GAGGTCCGCG TACCTTGCCT GTCTCCTACT CGCTCGTGGG AAAGACATCG
     1381 CGCCGTCGGA CGTGCAACGG AACGTCGATA AGATGAAGCG GTACATCCAC TTCGCGCCGG
     1441 GCAGTAAGGA CGAGTGGGCG GTCTCACTGT GCGGGATTCC CTCGCAGTCG CATCCACGGT
     1501 CCTTCTTGAC GCTGGCCAAC AACGGCTGCT TCCACAAGCC GATGCAGTGC CTGAAGGAGC
     1561 GATTTGTCGC CATGTACCGT GGGCGAGCAC ACCTCCATCA CTACCTGCTG AACGGGATGG
     1621 AGGCGGCCGA TTTCGAGGCC AGCTTGGAGA GCTTGTCAGC GTTGATCAGC GACTACAGGC
     1681 AAGTTGATGC TGCCGGCGCT GTCGCAGCCA ATGTGCGGGA TCGTGGATAC GTCGCCGTCG
     1741 GCGTGGTGAA GCCCAAGGTG AAGGCGGGCT GAcagtATCT TTCTAGAAGA TCTCCTACAA
     1801 TATTCTCAGC TGCCATGGAA AATCGATGTT CTTCTTTTAT TCTCTCAAGA TTTTCAGGCT
     1861 GTATATTAAA ACTTATATTA AGAACTATGC TAACCACCTC ATCAGGAACC GTTGTAGGTG
     1921 GCGTGGGTTT TCTTGGCAAT CGACTCTCAT GAAAACTACG AGCTAAATAT TCAATATGTT
     1981 CCTCTTGACC AACTTTATTC TGCATTTTTT TTGAACGAGG TTTAGAGCAA GCTTCAGGAA
     2041 ACTGAGACAG GAATTTTATT AAAAATTTAA ATTTTGAAGA AAGTTCAGGG TTAATAGCAT
     2101 CCATTTTTTG CTTTGCAAGT TCCTCAGCAT TCTTAACAAA AGACGTCTCT TTTGACATGT
     2161 TTAAAGTTTA AACCTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CATTATACGA
     2221 GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT
     2281 GCGTTGCGCT CACTGCCAAT TGCTTTCCAG TCGGGAAACC TGTCGTGCCA GCTGCATTAA
     2341 TGAATCGGCC AACGCGCGGG GAGAGGCGGT TTGCGTATTG GGCGCTCTTC CGCTTCCTCG
     2401 CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG CGGTATCAGC TCACTCAAAG
     2461 GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA
     2521 GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC
     2581 CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA
     2641 GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG
     2701 ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT
     2761 CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT
     2821 GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG
     2881 TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC
     2941 AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA CTACGGCTAC
     3001 ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA
     3061 GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC
     3121 AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG
     3181 GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT GAGATTATCA
     3241 AAAAGGATCT TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC AATCTAAAGT
     3301 ATATATGAGT AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA
     3361 GCGATCTGTC TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG
     3421 ATACGGGAGG GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA
     3481 CCGGCTCCAG ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT
     3541 CCTGCAACTT TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC TAGAGTAAGT
     3601 AGTTCGCCAG TTAATAGTTT GCGCAACGTT GTTGCCATTG CTACAGGCAT CGTGGTGTCA
     3661 CGCTCGTCGT TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA
     3721 TGATCCCCCA TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA
     3781 AGTAAGTTGG CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT
     3841 GTCATGCCAT CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA GTCATTCTGA
     3901 GAATAGTGTA TGCGGCGACC GAGTTGCTCT TGCCCGGCGT CAATACGGGA TAATACCGCG
     3961 CCACATAGCA GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC
     4021 TCAAGGATCT TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA
     4081 TCTTCAGCAT CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT
     4141 GCCGCAAAAA AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT CTTCCTTTTT
     4201 CAATATTATT GAAGCATTTA TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT
     4261 ATTTAGAAAA ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAC
     4321 GTCTAAGAAA CCATTATTAT CATGACATTA ACCTATAAAA ATAGGCGTAT CACGAGGCC
//