LOCUS       Exported                8696 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRRL-sffv-eGFP-cmv-hsGDNF, complete sequence.
ACCESSION   KJ697753
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8696)
  AUTHORS   Cordero Llana O.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-APR-2014) Physiology Anatomy and Genetics, University 
            of Oxford, South Parks Rd., Oxford, England OX1 3QX, United Kingdom
REFERENCE   2  (bases 1 to 8696)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8696
                     /organism="Cloning vector pRRL-sffv-eGFP-cmv-hsGDNF"
                     /mol_type="other DNA"
                     /db_xref="taxon:1615497"
     misc_feature    52..643
                     /label=WPRE
                     /note="WPRE"
     misc_feature    52..640
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(523..534)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    710..725
                     /label=cPPPT5
                     /note="cPPPT5"
     LTR             727..960
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    780..960
                     /label=HIV-LTR
                     /note="HIV-LTR"
     polyA_signal    1032..1153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1172..1307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(1340..1358)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1368..1384)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1529..1984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2013..2117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2118..2978
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /protein_id="AJO15941.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2118..2978
                     /gene="AmpR"
                     /label=AmpR
     rep_origin      3149..3737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4025..4046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4061..4091
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4099..4115
                     /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)."
     primer_bind     4123..4139
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4160..4178
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      4204..4433
                     /regulatory_class="promoter"
                     /note="rsv"
     misc_feature    4434..4614
                     /label=HIV-LTR
                     /note="HIV-LTR"
     LTR             4434..4614
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     primer_bind     4616..4766
     misc_feature    4658..4783
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5276..5509
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5277..5518
                     /label=rre
                     /note="rre"
     misc_feature    5999..6116
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6003..6018
                     /label=cPPPT3
                     /note="cPPPT3"
     regulatory      6155..6637
                     /regulatory_class="promoter"
                     /note="sffv"
     promoter        6233..6640
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      6656..6665
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6662..7381
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /protein_id="AJO15939.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      7449..8033
                     /regulatory_class="promoter"
                     /note="CMV"
     enhancer        7458..7761
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7762..7965
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             8060..8695
                     /codon_start=1
                     /product="GDNF"
                     /label=GDNF
                     /protein_id="AJO15940.1"
                     /translation="MKLWDVVAVCLVLLHTASAFPLPAGKRPPEAPAEDRSLGRRRAPF
                     ALSSDSNMPEDYPDQFDDVMDFIQATIKRLKRSPDKQMAVLPRRERNRQAAAANPENSR
                     GKGRRGQRGKNRGCVLTAIHLNVTDLGLGYETKEELIFRYCSGSCDAAETTYDKILKNL
                     SRNRRLVSDKVGQACCRPIAFDDDLSFLDDNLVYHILRKHSAKRCGCI"