back Return to this vector's summary.
ID   PHY163PLK  preliminary; circular DNA; SYN; 2519 BP.
XX
AC   D00375;
XX
DT   16-AUG-1990 (Rel. 5, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   Bacillus/E.coli plasmid vector pHY163PLK - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RP   1-2519
RC   pHY300-7PLK from pHY300PLK & pHY340
RC   pHY300-1PLK from pHY300-7PLK & linker
RC   pHY230PLK from pHY300-1PLK & linker
RC   pHY163PLK from pHY230PLK
RA   Ishiwa H., Shibahara H.;
RT   "New shuttle vectors for Escherichia coli and Bacillus subtilis.
RT   III. Nucleotide sequence analysis of tetracycline resistance gene
RT   of pAMalpha1 and ori-177";
RL   Jpn. J. Genet. 60:485-498(1985).
XX
RN   [2]
RC   pTHT15 from Bacillus stearothermophilus
RA   Furukawa K.;
RT   ;
RL   Unpublished (1985).
XX
CC   The tet alpha 1 is highly homologous to the tetracycline
CC   resistance genes of other Gram-positive bacteria, but completely
CC   different from the tetracycline resistance gene encoded by pBR322
CC   in Gram-negative bacteria.
CC   NM (pHY163PLK)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)(B.subtilis)(Gram-positive bacteria)(S.faecalis)
CC   CP ()
CC   FN (cloning)(shuttle)(transformation)
CC   SE ()
CC   PA (pAMalpha1)(ori-177)(p15A)(pHY300PLK)
CC   BR (pHY320PLK)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. S.faecalis, tet gene
FT                   -> pAMalpha1
FT                   1. pACYC177 3941bp, amp/kan gene
FT                   2. pAMalpha1 3000bp, tet gene
FT                   -> pHY460 7000bp
FT                   1. pHY460 remove between pAMalpha1 ori and amp 1100bp
FT                   spontaneous deletion
FT                   -> pHY385 5900bp [no XbaI, no AvaI, no BamHI]
FT                   1. pHY385 remove HaeII-HaeII 700bp, between amp & tet
FT                   \ pACYC177 HaeII = 1596 3026
FT                   -> pHY340 5200bp [unique HaeII]
FT                   1. pHY300PLK BanI-EcoRV 1623bp 4124..4870..877,
FT                   \ amp to tet
FT                   2. pHY340 remove BanI-EcoRV 2200bp, amp to tet/3000bp
FT                   -> pHY300-7PLK 4700bp [EcoRI between ori and amp]
FT                   1. pHY300-7PLK NcoI 4700bp, pAMalpha1 ori/2584..2584/
FT                   \ pHY300PLK 1027..1027 or 2584..2584
FT                   T4 DNA polymerase
FT                   XhoI linker 6bp ctcgag
FT                   -> pHY300-1PLK 4700bp
FT                   1. pHY300-1PLK XhoI 4700bp, pAMalpha1 ori
FT                   \ ?pACYC177 1953..1953
FT                   S1 nuclease
FT                   -> pHY230PLK 3303bp
FT                   1. pHY230PLK remove BanI-EcoRI 784bp, amp gene/2519bp
FT                   \ pHY300PLK 4124..4870..38
FT                   -> pHY163PLK 2519bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli miniplasmid p15A"
FT   variation       9..9
FT                   /note="c in pHY163PLK; cc in p15A"
FT   rep_origin      12..12
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   variation       30..30
FT                   /note="c in pHY163PLK; cc in p15A"
FT   misc_feature    414..414
FT                   /note="E. coli RNAI gene start site"
FT   misc_feature    complement(521..521)
FT                   /note="primer start site"
FT   -35_signal      883..888
FT                   /note="S35 Gram-positive bacteria -35 region"
FT   -10_signal      906..911
FT                   /note="S10 Gram-positive bacteria -10 region
FT                   (TATA box)"
FT   RBS             928..928
FT                   /note="RBS Gram-positive bacteria SD1"
FT   RBS             1032..1032
FT                   /note="RBS Gram-positive bacteria SD2"
FT   CDS             1044..2420
FT                   /note="ANT Gram-positive bacteria tetracycline
FT                   resistance gene alpha1 peptide, from pAMalpha1 and
FT                   ori-177"
FT   variation       2130..2131
FT                   /note="tc in pHY163PLK; ct in pTHT15"
FT   variation       2294..2294
FT                   /note="g in pHY163PLK; a in pTHT15"
FT   terminator      2427..2466
FT                   /note="TER Gram-positive bacteria transcriptional
FT                   terminator"
FT   misc_binding    2478..2519
FT                   /note="MCS"
XX
SQ   Sequence 2519 BP; 716 A; 458 C; 524 G; 821 T; 0 other;
     gggcaaagcg tttttccata ggctccgccc cctgacaagc atcacgaaat ctgacgctca
     aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     tccctcgtgc gctctcctgt tcctgccttt ccgtttaccg gtgtcattcc gctgttatgg
     ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     ccttcgaaaa acctccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     gaccaaaacg atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa
     actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     gttaccaatg cttaatcagt gaggcacaat tccttaagga acgtacagac gcttaaaagc
     ctttaaaaac gtttttaagg ggtttgtaga cggaaaaatt attttattaa agagtagttc
     aacaaacggg ccatattgtt gtataagtga tgaaatactg aatttaaaac ttagtttata
     tgtggtaaaa tgttttaatc aagtttagga ggaattaatt atgaagtgta atgaataatg
     aatgtaacag ggttcaatta aaagagggaa gcgtatcatt aaccctataa actacgtctg
     ccctcattat tggagggtga aatgtgaata catcctattc acaatcgaat ttacgacaca
     accaaatttt aatttggctt tgcattttat ctttttttag cgtattaaat gaaatggttt
     tgaacgtctc attacctgat attgcaaatg attttaataa accacctgcg agtacaaact
     gggtgaacac agcctttatg ttaacctttt ccattggaac agctgtatat ggaaagctat
     ctgatcaatt aggcatcaaa aggttactcc tatttggaat tataataaat tgtttcgggt
     cggtaattgg gtttgttggc cattctttct tttccttact tattatggct cgttttattc
     aaggggctgg tgcagctgca tttccagcac tcgtaatggt tgtagttgcg cgctatattc
     caaaggaaaa taggggtaaa gcatttggtc ttattggatc gatagtagcc atgggagaag
     gagtcggtcc agcgattggt ggaatgatag cccattatat tcattggtcc tatcttctac
     tcattcctat gataacaatt atcactgttc cgtttcttat gaaattatta aagaaagaag
     taaggataaa aggtcatttt gatatcaaag gaattatact aatgtctgta ggcattgtat
     tttttatgtt gtttacaaca tcatatagca tttcttttct tatcgttagc gtgctgtcat
     tcctgatatt tgtaaaacat atcaggaaag taacagatcc ttttgttgat cccggattag
     ggaaaaatat accttttatg attggagttc tttgtggggg aattatattt ggaacagtag
     cagggtttgt ctctatggtt ccttatatga tgaaagatgt tcaccagcta agtactgccg
     aaatcggaag tgtaattatt ttccctggaa caatgagtgt cattattttc ggctacattg
     gtgggatact tgttgataga agaggtcctt tatacgtgtt aaacatcgga gttacatttc
     tttctgttag ctttttaact gcttcctttc ttttagaaac aacatcatgg ttcatgacaa
     ttataatcgt atttgtttta ggtgggcttt cgttcaccaa aacagttata tcaacaattg
     tttcaagtag cttgaaacag caggaagctg gtgctggaat gagtttgctt aactttacca
     gctttttatc agagggaaca ggtattgcaa ttgtaggtgg tttattatcc atacccttac
     ttgatcaaag gttgttacct atggaagttg atcagtcaac ttatctgtat agtaatttgt
     tattactttt ttcaggaatc attgtcatta gttggctggt taccttgaat gtatataaac
     attctcaaag ggatttctaa atcgttaagg gatcaacttt gggagagagt tcaaaattga
     tccttttttt ataacaggaa ttcccgggga tccgtcgacc tgcagatctc tagaagctt
//