LOCUS       Exported                6825 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Retro and Lentiviral transduction of primary human germinal center B
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    phCMV-GALV-MTR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  AUTHORS   Caeser R, Di Re M, Krupka JA, Gao J, Lara-Chica M, Dias JML, Cooke 
            SL, Fenner R, Usheva Z, Runge HFP, Beer PA, Eldaly H, Pak HK, Park 
            CS, Vassiliou GS, Huntly BJP, Mupo A, Bashford-Rogers RJM, Hodson DJ
  TITLE     Genetic modification of primary human B cells to model high-grade 
            lymphoma.
  JOURNAL   Nat Commun. 2019 Oct 4;10(1):4543. doi: 10.1038/s41467-019-12494-x.
  PUBMED    31586074
REFERENCE   2  (bases 1 to 6825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-4"; volume: "10(1):4543. doi"; pages: " 
            10.1038/s41467-019-12494-x"
FEATURES             Location/Qualifiers
     source          1..6825
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        86..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     666..690
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          781..1353
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3519..3538)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3584..3639
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3638..3657)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3997..4013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3997..4013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4010..4032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4021..4037
                     /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(4045..4075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4090..4111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4228..4245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4399..4987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4479..4498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5158..6018)
                     /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     5781..5800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6019..6123)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6149..6604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6286..6307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6498..6517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6731..6753
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6745..6762
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6746..6762
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6772..6791
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6772..6790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"