LOCUS       Exported                5872 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pF151 pcDNA3.1(+)SOD1WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5872)
  AUTHORS   Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin 
            JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM
  TITLE     Modification of superoxide dismutase 1 (SOD1) properties by a GFP 
            tag--implications for research into amyotrophic lateral sclerosis 
            (ALS).
  JOURNAL   PLoS One. 2010 . 5(3):e9541.
  PUBMED    20221404
REFERENCE   2  (bases 1 to 5872)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One. 
            2010 . 5(3):e9541."
FEATURES             Location/Qualifiers
     source          1..5872
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     642..661
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        642..660
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1242..1259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1248..1472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1518..1946
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1605..1624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1815..1836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1955..1975)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1960..2289
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2140..2275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2202..2221
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2356..3150
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2410..2429)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3020..3039
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3324..3445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3361..3380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3415..3434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3494..3510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3494..3510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3507..3529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3518..3534
                     /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(3542..3572)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3587..3608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3725..3742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3896..4484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3976..3995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4655..5515)
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5695..5714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"