LOCUS       Exported                8314 bp ds-DNA     circular SYN 04-NOV-2021
DEFINITION  mammalian expression of rat GluA1.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV2-SEP-GluA1 (M1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8314)
  TITLE     Jeremy Henley Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8314)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..8314
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(3..22)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     249..269
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    293..915
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     697..716
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(3603..4313)
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3643..3663)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4258..4286
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(4496..4699)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4525..4545)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4700..5079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5240..5257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5240..5256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5249..5271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5397..5852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5534..5555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5746..5765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5879..5983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5984..6844
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6202..6221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7015..7603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7504..7523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7757..7774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7891..7912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7927..7957
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7965..7981
                     /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     7970..7992
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7989..8005
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7989..8005
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8019..8038
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8019..8037
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          8114..8210
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     promoter        complement(join(8249..8314,1..264))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8263..8314,1..84)
                     /label=SV40 ori
                     /note="SV40 origin of replication"