LOCUS       Exported                6846 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Control Lentiviral vector with ubiquitous PGK promoter, expresses 
            EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    PL-SIN-PGK-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS, 
            Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J
  TITLE     Isolation of human iPS cells using EOS lentiviral vectors to select 
            for pluripotency.
  JOURNAL   Nat Methods. 2009 May . 6(5):370-6.
  PUBMED    19404254
REFERENCE   2  (bases 1 to 6846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2009 May . 6(5):370-6."
FEATURES             Location/Qualifiers
     source          1..6846
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             785..1042
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    1693..1810
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2768..3267
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2803..2826)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3200..3219
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3281..3290
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3287..4006
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3287..4003
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3332..3353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3593..3612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3940..3961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4133..4366
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4403..4425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4414..4430
                     /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(4438..4468)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4483..4504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4621..4638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4872..4891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6173..6192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6411..6515)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(6552..6748)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6566..6701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6620..6639)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6819..6837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"