LOCUS       pJB553                  2880 bp    DNA     circular     03-APR-2015
DEFINITION   Lvl0 -98/5 SGP 1xTetApt-STOP.
ACCESSION   urn.local...iz-9e4hqd1
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..2880
                     /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"
     CDS             complement(146..469)
                     /gene="lacZalpha"
                     /codon_start=1
                     /product="beta-galactosidase alpha fragment"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLN
                     RLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTI
                     CSDAA"
     gene            complement(146..469)
                     /gene="lacZalpha"
     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"
     primer_bind     694..884
                     /standard_name="JB-553-SGP5-1xTetApt-STOP-gblock"
     misc_feature    701..704
                     /standard_name="BsaI Cut Site"
     misc_feature    702..804
                     /standard_name="98/5 SGP"
     primer_bind     719..740
                     /standard_name="TK-VEE-nsP4-seq-F"
     primer_bind     755..769
                     /standard_name="ApaI 15 bp Overhang"
     misc_feature    800..>804
                     /standard_name="Subgenomic RNA 5'UTR"
     misc_feature    805..851
                     /note="Geneious type: Aptamer"
                     /standard_name="5-1.31 Stem Loop"
     misc_feature    804^805
                     /note="Geneious type: Editing History Deletion"
                     /standard_name="CTACGACATAGTCTAGTCCGCCAAG"
     misc_feature    807..871
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="TGTATCCTTAACCATTTCTTTTATTGTATCCTTAACCATTTC
                     TTTT"
     CDS             836..868
                     /standard_name="ORF leader long"
     misc_feature    836..838
                     /standard_name="Start Codon"
     misc_feature    852..863
                     /standard_name="CAA repeats (aptamer spacer)"
     misc_feature    864..866
                     /standard_name="Spacer to prevent ATG"
     misc_feature    complement(874..877)
                     /standard_name="BsaI Cut Site"
     misc_feature    complement(879..884)
                     /standard_name="BsaI"
     primer_bind     complement(970..990)
                     /standard_name="TW_lvl0_seq_Rev1"
     rep_origin      complement(1002..1675)
     misc_RNA        complement(1061..1613)
                     /note="RNAII transcript (complementary strand)"
     rep_origin      complement(1061..1649)
     rep_origin      complement(1061..1594)
     primer_bind     complement(1100..1120)
                     /standard_name="TW_HDV-seq-R"
     primer_bind     1101..1122
                     /standard_name="pHAGE-A7550-7050"
     primer_bind     complement(1365..1389)
                     /standard_name="pHAGE-S7501-8000"
     primer_bind     complement(1409..1428)
                     /standard_name="RepO-R"
     -35_signal      1467..1472
                     /note="RNAI promoter (clockwise); TTGAAG"
     -10_signal      1489..1494
                     /note="RNAI promoter (clockwise); GCTACA"
     misc_RNA        1503..1610
                     /note="RNAI transcript"
     primer_bind     1612..1633
                     /standard_name="pHAGE-A7050-6550"
     -10_signal      complement(1623..1628)
                     /note="RNAII promoter (counter-clockwise); CGTAAT"
     -35_signal      complement(1644..1649)
                     /note="RNAII promoter (counter-clockwise); TTGAGA"
     CDS             complement(1820..2680)
                     /gene="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /codon_start=1
                     /product="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     gene            complement(1820..2680)
                     /gene="bla"
     primer_bind     complement(1869..1893)
                     /standard_name="pHAGE-S7001-7500"
     misc_feature    complement(1960..1965)
                     /standard_name="BsaI site"
     primer_bind     2121..2144
                     /standard_name="pHAGE-A6550-6050"
     primer_bind     complement(2389..2413)
                     /standard_name="pHAGE-S6501-7000"
     primer_bind     2443..2462
                     /standard_name="Amp-R; Quintara AmpR"
     primer_bind     2590..2613
                     /standard_name="pHAGE-A6050-5550"
     sig_peptide     complement(2612..2680)
                     /gene="bla"
                     /note="required for secretion to the periplasm; cleaved
                     off to form the mature beta-lactamase protein."
     promoter        complement(2681..2715)
                     /note="Beta Lactamase Promoter"
                     /standard_name="P-Bla"
     -10_signal      complement(2724..2729)
                     /note="bla promoter (counter-clockwise); GAGACA"
     -35_signal      complement(2745..2750)
                     /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 gacttccatc atagttatgg
      721 ccatgactac tctagctagc agtgttaaat cattcagcta cctgagaggg gcccctataa
      781 ctctctacgg ctaacctgaa tggaatccag gcagagaaag gtcgatacgg acggaatgtg
      841 gtggcctgga tcaacaacaa caacactgta agccacctga gaccgcttcc tcgctcactg
      901 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      961 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1501 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1921 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2401 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2821 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
