LOCUS       pPtGG-1      3742 bp    DNA     circular UNA 18-OCT-2007
DEFINITION  Cloning vector pBR322, complete sequence.
KEYWORDS    .
SOURCE      Cloning vector pBR322 (unknown)
  ORGANISM  Cloning vector pBR322
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 4361)
  AUTHORS   New England Biolabs.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2007) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA
COMMENT     1.  pBR322 beta-lactamase (bla, conferring ampicillin resistance)
            is transcribed in vitro and in vivo both from its natural promoter
            (P3) and from promoter P1, a pSC101 promoter artificially
            juxtaposed with the bla sequence during construction of the pBR322
            plasmid.  (Ref: Brosius et al, J Biol Chem 257, 9205-9210 (1982)).
            2.  Corrects sequence error in tet gene deriving from the original
            pSC101 sequence, present in GenBank entry (see accession J01749)
            and NEB sequences dated 6/6/02 and earlier, based on sequencing of
            pBR322 and pACYC184 at NEB in 2007 (Mathieu-Sheltry, V. and Cantin,
            E., unpublished).  Correction is C1134T, representing a Thr350Ile
            correction in the expressed protein.
FEATURES             Location/Qualifiers
     source          join(3369..3742,1..3368)
                     /organism="Cloning vector pBR322"
                     /mol_type="other DNA"
     CDS             join(3454..3742,1..902)
                     /gene="tet"
                     /note="tetR (confers resistance to tetracycline)"
                     /codon_start=1
                     /product="tetracycline efflux protein, class C"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASH
                     YGVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAG
                     RIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLH
                     APFLAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTV
                     FFIMQLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGE
                     KQAIIAGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQG
                     QLQGSLAALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSR
                     ATST"
     gene            join(3454..3742,1..902)
                     /gene="tet"
     RBS             1531..1535
                     /note="rop RBS"
     CDS             1541..1732
                     /gene="rop"
                     /exception="alternative start codon"
                     /codon_start=1
                     /transl_table=11
                     /product="ROP protein"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
     gene            1541..1732
                     /gene="rop"
     CDS             1836..1976
                     /cds_type=ORF
                     /label=OriT
     Site            join(1843..1851,1846^1847)
                     /site_type="restriction site"
                     /note="Name: Tth111I"
                     /note="Pattern: gacnnngtc"
                     /note="Number_of_matches: 1"
                     /note="cut_0_on_positive_strand: 2220^2221"
                     /note="cut_0_on_negative_strand: 2221^2222"
                     /note="inhibited_by: 5': N6-methyladenosine"
                     /note="star_activity: yes"
                     /note="site_type: other"
                     /note="restriction site"
                     /label=Tth111I
     misc_RNA        complement(2160..2712)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(2160..2748)
                     /note="pMB1 origin of replication (counter-clockwise)
                     (RNAII -35 to RNA/DNA switch point)"
     -35_signal      2566..2571
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      2588..2593
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        2602..2709
                     /note="RNAI transcript"
     -10_signal      complement(2722..2727)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(2743..2748)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     primer          complement(2782..2829)
                     /label=3'hpRNA-R-VECTOR
     primer          2810..2855
                     /label=5'hpRNA-F-OH
     LTR             complement(2878..2977)
                     /label=attL1
     primer          complement(2940..2977)
                     /label=PtRNAi-40
     Site            2998..3046
                     /site_type="restriction site"
                     /label=MCS1
     intron          3047..3138
                     /label="white intron/I-SceI"
     Site            3139..3178
                     /site_type="restriction site"
                     /label=MCS2
     primer          3195..3228
                     /label=PtRNAi-39
     LTR             3195..3294
                     /label=attL2
     primer          complement(3295..3341)
                     /label=3'hpRNA-R-OH
     primer          3322..3366
                     /label=5'hpRNA-F-VECTOR
     -35_signal      3378..3383
                     /note="tet (P2) promoter (clockwise); TTGACA"
     -10_signal      3401..3406
                     /note="tet (P2) promoter (clockwise); TTTAAT; transcript
                     start 45"
     -10_signal      complement(3412..3417)
                     /note="P1 promoter (counter-clockwise); TAAACT"
     -35_signal      complement(3432..3443)
                     /note="P1 promoter -35 sequence region; transcript starts
                     36, 37 (complementary strand)"
ORIGIN
        1 GGATCCTCTA CGCCGGACGC ATCGTGGCCG GCATCACCGG CGCCACAGGT GCGGTTGCTG
       61 GCGCCTATAT CGCCGACATC ACCGATGGGG AAGATCGGGC TCGCCACTTC GGGCTCATGA
      121 GCGCTTGTTT CGGCGTGGGT ATGGTGGCAG GCCCCGTGGC CGGGGGACTG TTGGGCGCCA
      181 TCTCCTTGCA TGCACCATTC CTTGCGGCGG CGGTGCTCAA CGGCCTCAAC CTACTACTGG
      241 GCTGCTTCCT AATGCAGGAG TCGCATAAGG GAGAGCGTCG ACCGATGCCC TTGAGAGCCT
      301 TCAACCCAGT CAGCTCCTTC CGGTGGGCGC GGGGCATGAC TATCGTCGCC GCACTTATGA
      361 CTGTCTTCTT TATCATGCAA CTCGTAGGAC AGGTGCCGGC AGCGCTCTGG GTCATTTTCG
      421 GCGAGGACCG CTTTCGCTGG AGCGCGACGA TGATCGGCCT GTCGCTTGCG GTATTCGGAA
      481 TCTTGCACGC CCTCGCTCAA GCCTTCGTCA CTGGTCCCGC CACCAAACGT TTCGGCGAGA
      541 AGCAGGCCAT TATCGCCGGC ATGGCGGCCG ACGCGCTGGG CTACGTCTTG CTGGCGTTCG
      601 CGACGCGAGG CTGGATGGCC TTCCCCATTA TGATTCTTCT CGCTTCCGGC GGCATCGGGA
      661 TGCCCGCGTT GCAGGCCATG CTGTCCAGGC AGGTAGATGA CGACCATCAG GGACAGCTTC
      721 AAGGATCGCT CGCGGCTCTT ACCAGCCTAA CTTCGATCAT TGGACCGCTG ATCGTCACGG
      781 CGATTTATGC CGCCTCGGCG AGCACATGGA ACGGGTTGGC ATGGATTGTA GGCGCCGCCC
      841 TATACCTTGT CTGCCTCCCC GCGTTGCGTC GCGGTGCATG GAGCCGGGCC ACCTCGACCT
      901 GAATGGAAGC CGGCGGCACC TCGCTAACGG ATTCACCACT CCAAGAATTG GAGCCAATCA
      961 ATTCTTGCGG AGAACTGTGA ATGCGCAAAC CAACCCTTGG CAGAACATAT CCATCGCGTC
     1021 CGCCATCTCC AGCAGCCGCA CGCGGCGCAT CTCGGGCAGC GTTGGGTCCT GGCCACGGGT
     1081 GCGCATGATC GTGCTCCTGT CGTTGAGGAC CCGGCTAGGC TGGCGGGGTT GCCTTACTGG
     1141 TTAGCAGAAT GAATCACCGA TACGCGAGCG AACGTGAAGC GACTGCTGCT GCAAAACGTC
     1201 TGCGACCTGA GCAACAACAT GAATGGTCTT CGGTTTCCGT GTTTCGTAAA GTCTGGAAAC
     1261 GCGGAAGTCA GCGCCCTGCA CCATTATGTT CCGGATCTGC ATCGCAGGAT GCTGCTGGCT
     1321 ACCCTGTGGA ACACCTACAT CTGTATTAAC GAAGCGCTGG CATTGACCCT GAGTGATTTT
     1381 TCTCTGGTCC CGCCGCATCC ATACCGCCAG TTGTTTACCC TCACAACGTT CCAGTAACCG
     1441 GGCATGTTCA TCATCAGTAA CCCGTATCGT GAGCATCCTC TCTCGTTTCA TCGGTATCAT
     1501 TACCCCCATG AACAGAAATC CCCCTTACAC GGAGGCATCA GTGACCAAAC AGGAAAAAAC
     1561 CGCCCTTAAC ATGGCCCGCT TTATCAGAAG CCAGACATTA ACGCTTCTGG AGAAACTCAA
     1621 CGAGCTGGAC GCGGATGAAC AGGCAGACAT CTGTGAATCG CTTCACGACC ACGCTGATGA
     1681 GCTTTACCGC AGCTGCCTCG CGCGTTTCGG TGATGACGGT GAAAACCTCT GACACATGCA
     1741 GCTCCCGGAG ACGGTCACAG CTTGTCTGTA AGCGGATGCC GGGAGCAGAC AAGCCCGTCA
     1801 GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG GGGCGCAGCC ATGACCCAGT CACGTAGCGA
     1861 TAGCGGAGTG TATACTGGCT TAACTATGCG GCATCAGAGC AGATTGTACT GAGAGTGCAC
     1921 CATATGCGGT GTGAAATACC GCACAGATGC GTAAGGAGAA AATACCGCAT CAGGCGCTCT
     1981 TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA
     2041 GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC
     2101 ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT
     2161 TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG
     2221 CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC
     2281 TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC
     2341 GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC
     2401 AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC
     2461 TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT
     2521 AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT
     2581 AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC
     2641 TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT
     2701 TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG
     2761 ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG CGCAACGTTG
     2821 TTGCCATTGT CCCAGTCACG ACGTTGTAAA ACGACGGCCA GTCTTAAGCT CGGGCCCCAA
     2881 ATAATGATTT TATTTTGACT GATAGTGACC TGTTCGTTGC AACAAATTGA TGAGCAATGC
     2941 TTTTTTATAA TGCCAACTTT GTACAAAAAA GCAGGCTCCG CGGCCGCCCC CTTcaccTGG
     3001 GAATTCGTTA ACAGATCTGa GaCCGCGGCT CGAGACTAGT GCCTAGGTGA GTTTCTATTC
     3061 GCAGTCGGCT GATCTGTGTG AAAtagggat aacagggtaa tTCTTAATGA AGGGTCCAAT
     3121 TACCAATTTG AAACTCAGCT AGCTCTAGAA TATCAATTGG GATCCggtct catcgacaAA
     3181 GGGTGGGCGC GCCGACCCAG CTTTCTTGTA CAAAGTTGGC ATTATAAGAA AGCATTGCTT
     3241 ATCAATTTGT TGCAACGAAC AGGTCACTAT CAGTCAAAAT AAAATCATTA TTTGCCATCC
     3301 AGCTGATATC CCCTATAGTG AAGGGTCAGT GCTGCAACAT TATCACGAGG CCCTTTCGTC
     3361 TTCAAGAATT CTCATGTTTG ACAGCTTATC ATCGATAAGC TTTAATGCGG TAGTTTATCA
     3421 CAGTTAAATT GCTAACGCAG TCAGGCACCG TGTATGAAAT CTAACAATGC GCTCATCGTC
     3481 ATCCTCGGCA CCGTCACCCT GGATGCTGTA GGCATAGGCT TGGTTATGCC GGTACTGCCG
     3541 GGCCTCTTGC GGGATATCGT CCATTCCGAC AGCATCGCCA GTCACTATGG CGTGCTGCTA
     3601 GCGCTATATG CGTTGATGCA ATTTCTATGC GCACCCGTTC TCGGAGCACT GTCCGACCGC
     3661 TTTGGCCGCC GCCCAGTCCT GCTCGCTTCG CTACTTGGAG CCACTATCGA CTACGCGATC
     3721 ATGGCGACCA CACCCGTCCT GT
//
