LOCUS       Exported                6815 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Keratinocyte expression vector phPK14H, complete sequence.
ACCESSION   DQ898181
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6815)
  AUTHORS   Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
  TITLE     A recombinant plasmid for specific expression of recombinant protein
            in human kertinocytes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6815)
  AUTHORS   Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
  TITLE     Keratinocyte specific-expression vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6815)
  AUTHORS   Hosseini SJ, Zomorodipour A, Jalal R, Sabouni F.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2006) Molecular Genetics, The National Institute 
            for Genetic Engineering and Biotechnology, Blvd. Pjoohesh. km-17 
            Tehran-karaj Highway, Tehran, Tehran 14155-6343, Iran
REFERENCE   4  (bases 1 to 6815)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6815
                     /organism="Keratinocyte expression vector phPK14H"
                     /mol_type="other DNA"
                     /db_xref="taxon:402237"
     regulatory      18..2557
                     /regulatory_class="promoter"
                     /note="Homo sapiens promoter for keratin 14 (PhK14)"
     CDS             complement(260..277)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2258..2363
                     /label=multiple cloning site (MCS)
                     /note="multiple cloning site (MCS)"
     promoter        complement(2367..2385)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2411..2635
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2618..3287
                     /label=BGH polyA site
                     /note="BGH polyA site"
     rep_origin      2681..3109
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3123..3453
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     regulatory      3159..3484
                     /regulatory_class="promoter"
                     /note="SV40 promoter"
     rep_origin      3304..3439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      3353..3438
                     /label=SV40 Ori
                     /note="SV40 Ori"
     CDS             3520..4314
                     /codon_start=1
                     /gene="Neo"
                     /product="Neo"
                     /label=Neo
                     /note="neomycin resistance protein"
                     /protein_id="ABI64163.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3520..4314
                     /gene="Neo"
                     /label=Neo
     misc_feature    4369..4741
                     /label=SV40 polyA site
                     /note="SV40 polyA site"
     polyA_signal    4488..4609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4658..4674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4682..4698
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4706..4736)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4751..4772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      5001..5674
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
     rep_origin      complement(5060..5648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5819..6679)
                     /codon_start=1
                     /gene="amp"
                     /gene_synonym="bla"
                     /product="Amp"
                     /label=amp
                     /note="ampicillin resistance protein; beta-lactamase"
                     /protein_id="ABI64164.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5819..6679)
                     /gene="amp"
                     /gene_synonym="bla"
                     /label=amp
     promoter        complement(6680..6784)
                     /gene="bla"
                     /label=AmpR promoter