LOCUS       pJB332                  4810 bp    DNA     circular     03-OCT-2014
DEFINITION  Lvl0 TetR-Gly9-DDX6 (contains SapI).
ACCESSION   urn.local...kq-9e4hqeg
KEYWORDS    .
SOURCE      Cloning vector pUC19
  ORGANISM  Cloning vector pUC19
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 2686)
  AUTHORS   New, England and Biolabs.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2007) Research Department, New England Biolabs,
            240 County Road, Ipswich, MA 01938, USA COMMENT See also GenBank
            accession L09137.
FEATURES             Location/Qualifiers
     source          1..4810
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
     gene            55..655
                     /standard_name="lacZ PCR Product from pUC19"
     primer_bind     55..75
                     /standard_name="TK_DI_lacZ_Fwd_1"
     gene            complement(146..469)
                     /gene="lacZalpha"
     CDS             complement(146..469)
                     /gene="lacZalpha"
                     /codon_start=1
                     /product="beta-galactosidase alpha fragment"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLN
                     RLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTI
                     CSDAA"
     primer_bind     364..386
                     /standard_name="M13/pUC Forward (Quintara)"
     primer_bind     378..395
                     /standard_name="M13 (-21) Forward (Quintara)"
     misc_feature    396..452
                     /gene="lacZalpha"
                     /note="multiple cloning site (EcoRI-HindIII)"
                     /note="Geneious type: polylinker"
     primer_bind     complement(465..481)
                     /standard_name="M13 Reverse (Quintara)"
     -10_signal      complement(514..519)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TATGTT"
     -35_signal      complement(538..543)
                     /note="Plac promoter (drives lacZalpha;
                     counter-clockwise); TTTACA"
     protein_bind    complement(563..575)
                     /note="CAP protein binding site"
                     /bound_moiety="catabolite activator protein (CAP)"
     primer_bind     578..598
                     /standard_name="TW_lvl0_seq_Fwd1"
     misc_feature    679..693
                     /standard_name="15 bp SapI Overhang"
     misc_feature    694..699
                     /standard_name="BsaI"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     source          <705..>1325
                     /organism="Cloning vector pUC19"
                     /mol_type="other DNA"
     CDS             705..1325
                     /standard_name="TetR"
     misc_feature    969..975
                     /standard_name="SapI Site"
     CDS             1326..1352
                     /standard_name="Gly 9 Linker"
     CDS             1353..2801
                     /standard_name="DDX6 (transcript 3)"
     misc_feature    complement(2700..2706)
                     /standard_name="SapI Site"
     misc_feature    complement(2804..2807)
                     /standard_name="BsaI Cut Site"
     primer_bind     complement(<2805..2814)
                     /standard_name="JB-280-Myr-Akt1-HA-PCR-R"
     misc_feature    complement(2809..2814)
                     /standard_name="BsaI"
     primer_bind     complement(2900..2920)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(2932..3605)
     misc_RNA        complement(2991..3543)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(2991..3579)
     rep_origin      complement(2991..3524)
     primer_bind     complement(3030..3050)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     3031..3052
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(3295..3319)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(3339..3358)
                     /standard_name="RepO-R"
     -35_signal      3397..3402
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      3419..3424
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        3433..3540
                     /note="RNAI transcript"
     primer_bind     3542..3563
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(3553..3558)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(3574..3579)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     gene            complement(3750..4610)
                     /gene="bla"
     CDS             complement(3750..4610)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     primer_bind     complement(3799..3823)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(3890..3895)
                     /standard_name="BsaI site"
     primer_bind     4051..4074
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(4319..4343)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     4373..4392
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     4520..4543
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(4542..4610)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(4611..4645)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(4654..4659)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(4675..4680)
                     /note="bla promoter (counter-clockwise); TTCAAA"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      661 agaggcggtt tgcgtattgg gcgctcttcc gctggtctcc caccatgtca agactcgaca
      721 agagcaaggt gattaacagt gcactggaac ttctcaatga agttgggatc gaggggctga
      781 ctactagaaa actcgcacag aaactggggg ttgagcagcc caccttgtac tggcacgtta
      841 aaaacaaaag ggccctgctg gatgctctgg ccatcgagat gctggatagg catcataccc
      901 acttctgccc tctggaagga gaatcctggc aggatttcct tagaaacaac gccaagtcct
      961 ttcgctgtgc tcttcttagc caccgggatg gtgctaaagt ccatctcggc acacgaccaa
     1021 ctgagaagca gtacgaaact ctcgagaacc agctggcctt tctctgtcaa cagggctttt
     1081 ctcttgaaaa cgccctgtac gcactgagtg cagttgggca ctttacactc ggatgtgttc
     1141 tggaggacca agaacatcag gtggcaaagg aagagaggga gacccctacg actgactcca
     1201 tgccccctct cttgaggcag gcaatagaat tgttcgacca tcagggcgca gaacccgcct
     1261 ttctgtttgg gctggaactg attatctgcg gtcttgagaa acagctgaag tgcgagtccg
     1321 ggagcggcgg tggcggtggc ggtggcggtg gtatgagcac agctcgcacc gagaacccgg
     1381 tgattatggg cctgtccagc cagaacggac agctcagagg gcctgtaaag gcttcagcag
     1441 gccccggcgg aggcggcaca caaccacaac cacagcttaa tcagcttaag aatactagca
     1501 ctattaataa cggaacaccg cagcaggccc aaagcatggc tgccacaatt aaacccggag
     1561 atgactggaa gaagaccctg aagctccctc caaaagatct caggattaaa actagcgatg
     1621 ttacttcaac aaagggaaat gagttcgaag actactgtct gaagcgagag ttgctgatgg
     1681 ggattttcga aatgggctgg gagaagccct ctcctattca agaagagagc atccccatcg
     1741 ctctgtccgg gagggacatc cttgccaggg ctaaaaatgg gaccggaaaa tcaggagctt
     1801 acttgatccc actccttgaa aggcttgatc tcaagaagga caacatccaa gctatggtta
     1861 tcgtgccaac tagagaactc gccctccagg tcagccagat ttgcatccag gtgagtaagc
     1921 acatgggcgg agctaaggtg atggctacaa ctggagggac taacctgcga gacgacataa
     1981 tgagacttga tgacacagtc catgtggtca tcgctacacc tgggaggatt ctggatctga
     2041 tcaaaaaagg agtggcaaag gtggatcatg tgcagatgat agtcttggac gaggccgaca
     2101 aactgctgag ccaagacttt gtgcagatca tggaggatat catcttgaca ctccccaaga
     2161 accgacagat tctgctgtac tccgcaacat ttcctctttc cgttcagaaa ttcatgaact
     2221 cacatctcca gaaaccttat gagatcaatt tgatggaaga actgacactg aagggcgtga
     2281 cccagtatta tgcctacgtt actgagaggc aaaaggtcca ctgcctgaat actctcttct
     2341 ccaggctcca gatcaaccag tctatcatct tttgcaatag ctcccagcga gtcgagctgc
     2401 tggctaagaa gatctcacag cttggatatt cctgtttcta catccatgct aagatgagac
     2461 aagagcacag aaaccgcgtc tttcatgatt tccggaacgg actctgtcgc aacctggttt
     2521 gcacagatct ttttactaga ggcatcgata tccaagcagt gaacgtggtt atcaacttcg
     2581 actttcccaa actcgccgag acttatcttc atagaattgg ccgatccggt aggtttgggc
     2641 acctggggct cgccatcaat ctcattacgt atgatgatag gttcaacctc aagtcaatag
     2701 aagagcagtt ggggaccgag atcaaaccaa tcccgagcaa tattgacaaa tcactctatg
     2761 tggccgaata ccattcagag cctgccgagg atgagaagcc ttaataatga gaccgcttcc
     2821 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2881 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2941 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3001 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3061 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4741 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4801 ccctttcgtc
//
