LOCUS       Exported                4801 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  recombinant AAV vector packaging self-complementary GFP under the 
            CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pscAAV-CAG-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4801)
  AUTHORS   Pekrun K, De Alencastro G, Luo QJ, Liu J, Kim Y, Nygaard S, Galivo 
            F, Zhang F, Song R, Tiffany MR, Xu J, Hebrok M, Grompe M, Kay MA
  TITLE     Using a barcoded AAV capsid library to select for clinically 
            relevant gene therapy vectors.
  JOURNAL   JCI Insight. 2019 Nov 14;4(22). pii: 131610. doi: 
            10.1172/jci.insight.131610.
  PUBMED    31723052
REFERENCE   2  (bases 1 to 4801)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "JCI 
            Insight."; date: "2019-11-14"; volume: "4(22). pii: 131610. doi"; 
            pages: " 10.1172/jci.insight.131610"
FEATURES             Location/Qualifiers
     source          1..4801
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        417..696
                     /label=chicken beta-actin promoter
     primer_bind     975..997
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1059..1078
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1118..1127
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1124..1843
                     /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             1124..1840
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1169..1190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1430..1449)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1777..1798
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1887..1906
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2156..2173)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2156..2172)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2165..2187)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2381..2400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2500..2522
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2560..2578)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2646..2750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2751..3611
                     /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(2969..2988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3782..4370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4271..4290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4524..4541
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4658..4679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4694..4724
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4732..4748
                     /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     4737..4759
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4756..4772
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4756..4772
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"