LOCUS       Exported                5992 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  express Assembly-activating protein (AAP) of adeno-associated virus 
            serotype 2 (AAV2).
ACCESSION   .
VERSION     .
KEYWORDS    pAAP2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  AUTHORS   Zinn E, Pacouret S, Khaychuk V, Turunen HT, Carvalho LS, 
            Andres-Mateos E, Shah S, Shelke R, Maurer AC, Plovie E, Xiao R, 
            Vandenberghe LH
  TITLE     In Silico Reconstruction of the Viral Evolutionary Lineage Yields a 
            Potent Gene Therapy Vector.
  JOURNAL   Cell Rep. 2015 Jul 28. pii: S2211-1247(15)00759-7. doi: 
            10.1016/j.celrep.2015.07.019.
  PUBMED    26235624
REFERENCE   2  (bases 1 to 5992)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep. 
            2015 Jul 28. pii: S2211-1247(15)00759-7. doi: 
            10.1016/j.celrep.2015.07.019."
FEATURES             Location/Qualifiers
     source          1..5992
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1586..1603)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1862..2290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1949..1968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2159..2180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2299..2319)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2304..2633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2484..2619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2546..2565
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2700..3494
                     /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(2754..2773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3364..3383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3668..3789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3705..3724)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3759..3778
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3838..3854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3838..3854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3851..3873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3862..3878
                     /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(3886..3916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3931..3952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4069..4086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4240..4825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4320..4339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4996..5856)
                     /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     5619..5638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5857..5961)
                     /gene="bla"
                     /label=AmpR promoter