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ID   PHGN3      preliminary; circular DNA; SYN; 4283 BP.
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AC   M60387; ATCC69051;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   Vertebrate/E.coli plasmid vector pHGN3 - incomplete, RPV virulence.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-947
RC   RPmuhr23 from rabbit pox virus mutant with HA gene
RC   pPvuII from pUC series
RC   pHGN1 from pUC9 & RPmuhr23
RC   pHGN2 from pHGN1 & pPvuII
RC   pHGN3, pHGN3.1 from pHGN2 & pBluescript KS+
RA   Bloom D.C., Edwards K.M., Hager C., Moyer R.W.;
RT   "Identification and characterization of two nonessential regions of
RT   the rabbitpox virus genome involved in virulence";
RL   J. Virol. 65:1530-1542(1991).
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RN   [2]
RC   pHGN3.1 from pHGN2 & pBluescript KS+
RA   ;
RT   ;
RL   Unpublished (1993).
RL   U.S. Patent No. 5,212,057 dated May 18, 1993.
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CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--4.2; EcoRI--4.2; SalI/HindIII--3.2, 1.0. (ATCC staff)
CC   Shuttle vector component of a virulence tester system to be used with
CC   the avirulent rabbit poxvirus deletion mutant RPmuhr23HA+
CC   (ATCC VR-2379). [2]
CC   DNA cloned into the MCS is integrated into VR-2379 by homologous
CC   recombination, possibly altering the host range and virulence of the
CC   virus.[2]
CC   The hemagglutinin insert is a 1.8 kb 5'SalI/3'HindIII fragment
CC   containing approximately 443 bp 5' flanking sequence, 988 bp coding
CC   sequence interrupted by a polylinker, and 408 bp 3' flanking sequence.
CC   [2]
CC   The gene is interrupted at the AccI site (nt 43 of the coding region)
CC   by a 47 bp MCS from pUC9. [2]
CC   Constructed by deleting lacZ and the polylinker from pUC9 by a HaeII
CC   digest. [2]
CC   The order of the major features in the plasmid is:  pUC9 delta lacZ -
CC   KpnI - 5' end of insert - PstI/MCS/EcoRI - 3' end of insert - KpnI.[2]
CC   Recombinants can be identified by inactivation of the hemagglutinin
CC   gene (HA-). [2]
CC   Medium is 1227 LB plus ampicillin.
CC   RPV = rabbit pox virus, vaccinia virus
CC   NCBI gi: 335806
CC   NM (pHGN3)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(vertebrate cells)
CC   HO (chicken embryo fibroblasts virus-infected)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. rabbit pox virus mutant
FT                   -> RPmuhr23 [30000bp removed from left end]
FT                   1. pUC18 remove PvuII-PvuII, lacZ gene
FT                   -> pPvuII
FT                   1. pUC9 remove HindIII-EcoRI, MCS
FT                   2. vaccinia virus HindIII-EcoRI 7800bp, HA gene
FT                   -> pHGN1
FT                   1. pHGN1 HindIII-SalI 1800bp, HA gene
FT                   T4 DNA polymerase fill in:T4 DNA polymerase fill in
FT                   2. pPvuII PvuII
FT                   -> pHGN2
FT                   1. pHGN2 NruI
FT                   2. pBluescript KS+ KpnI-SacI, MCS
FT                   T4 DNA polymerase fill in:T4 DNA polymerase fill in
FT                   -> pHGN3 4283bp"
FT   misc_binding    0..0
FT                   /note="MCS unique PstI-AccI-BamHI-SmaI-XmaI-EcoRI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             156..914
FT                   /note="GEN vaccinia virus gene involved in RPV
FT                   virulence; NCBI gi: 335807"
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SQ   Sequence 947 BP; 324 A; 143 C; 167 G; 313 T; 0 other;
     aattgtgtaa agaaaaacta attagattct cccacatttt tgttaacatt acactaacta
     attggtaaaa ttgatagaat aattatgtgt atataagata gatttcctat tgtcttactc
     attgcatcgt actattgtta catattttga gtaagatggt ttacaaatta gtcttactct
     tttgtatcgc atctctcggg tattcggtag aatacaagaa tactatatgt cctcctcgac
     aagattaccg gtattggtac tttgccgccg tactcactat tggtgtaaat tatgacatta
     attctactat tattggtgag tgtcatatga gtgaaagcta tatcgacaga aatgctaaca
     tagtgttgac tggatacgga ctagaaataa acatgaccat catggatacg gatcagagat
     ttgtggcagc ggctgagggt gttggtaaag acaataaact atctgttctg ttgtttacca
     ctcagcggct agataaagtt catcacaata ttagtgtgac aataacatgt atggaaatga
     attgtggaac cacaaaatac gatagcgatc ttccggaatc atttcataaa tcatcatcgt
     gtgatataac tataaatgga tcatgtgtga catgtgttaa cttagagact gatccaacaa
     agattaatcc ccattaccta caccccaagg ataaatatct ttatcataat tctgagtata
     gcatgcgtgg tagttatggc gtgacattta tagatgaact aaaccagtgc cttcttgaca
     taaaagaagt tatgatattt gttatagaga gtaaattgca aatatacaga aatagaaata
     gaaatattaa tttttatact acaatggaca acattatgtg gccagtatta tctatctctt
     tatgccaaat ataatagtaa aaatttagat gtatttagga atatgct
//