LOCUS       .                       2974 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    1..51
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     misc_feature    2931..2974
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     CDS             complement(1551..2411)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     protein_bind    411..427
                     /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)."
                     /note="color: #31849b"
                     /label="lac operator"
     promoter        2908..2926
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     promoter        complement(435..465)
                     /note="E. coli lac promoter with an 'up' mutation"
                     /note="color: #ffffff"
                     /label="lac UV5 promoter"
     promoter        complement(2412..2516)
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     rep_origin      complement(792..1380)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     CDS             complement(1..382)
                     /codon_start=1
                     /function=""
                     /gene="eco47IR"
                     /note="cleaves double-stranded DNA at GGWCC sites in the
                     absence of protective methylation"
                     /note="color: #993366"
                     /product="mutant version of the Eco47I restriction
                     endonuclease"
                     /protein_id=""
                     /transl_table=1
                     /translation="MSKETSFVKNAEELAKQKMDAINPELSSKFKFLIKFLSQFPEACS
                     KPRSKKMQNKVGQEEHIEYLARSFHESRLPRKPTPPTTVPDEVVSIVLNISFNIQPENL
                     ERIKEEHRFSMAAENIVGDLLER"
                     /label="Eco47I/T7"
     CDS             complement(2619..2974)
                     /codon_start=1
                     /function=""
                     /gene="eco47IR"
                     /note="cleaves double-stranded DNA at GGWCC sites in the
                     absence of protective methylation"
                     /note="color: #993366"
                     /product="mutant version of the Eco47I restriction
                     endonuclease"
                     /protein_id=""
                     /transl_table=1
                     /translation="LAEKLEPSGRSGGRSPYSESYYAGWIWCSGTSVKAVDFIHYDEKN
                     NEWNLLQVKNRDNTENSSSSKIRDNTTIKKWFRTYSQRDATNWDNFPDEVSSKNLNEED
                     FRAFVKNYLAKII*"
                     /label="Eco47I/T7"
ORIGIN
        1 atctttctag aagatctcct acaatattct cagctgccat ggaaaatcga tgttcttctt
       61 ttattctctc aagattttca ggctgtatat taaaacttat attaagaact atgctaacca
      121 cctcatcagg aaccgttgta ggtggcgtgg gttttcttgg caatcgactc tcatgaaaac
      181 tacgagctaa atattcaata tgttcctctt gaccaacttt attctgcatt ttttttgaac
      241 gaggtttaga gcaagcttca ggaaactgag acaggaattt tattaaaaat ttaaattttg
      301 aagaaagttc agggttaata gcatccattt tttgctttgc aagttcctca gcattcttaa
      361 caaaagacgt ctcttttgac atgtttaaag tttaaacctc ctgtgtgaaa ttgttatccg
      421 ctcacaattc cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
      481 tgagtgagct aactcacatt aattgcgttg cgctcactgc caattgcttt ccagtcggga
      541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      601 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      961 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1201 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1381 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1441 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1501 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1561 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1621 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1681 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1741 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1801 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1861 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1921 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1981 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2041 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2101 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2161 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2221 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2281 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2341 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2401 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2461 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2521 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     2581 aaaaataggc gtatcacgag gccgcccctg cagccgaatt atattatttt tgccaaataa
     2641 tttttaacaa aagctctgaa gtcttcttca tttaaattct tagatgatac ttcatctgga
     2701 aaattgtccc aattagtagc atcacgctgt gagtaagttc taaaccattt ttttattgtt
     2761 gtattatctc taatcttact actcgatgag ttttcggtat tatctctatt tttaacttgg
     2821 agcaggttcc attcattgtt tttttcatca tagtgaataa aatcaactgc tttaacactt
     2881 gtgcctgaac accatatcca tccggcgtaa tacgactcac tatagggaga gcggccgcca
     2941 gatcttccgg atggctcgag tttttcagca agat
//
