LOCUS       Exported                7385 bp ds-DNA     circular SYN 17-JUN-2021
DEFINITION  Human codon-optimized T7 polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    T7opt in pCAGGS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Yun T, Park A, Hill TE, Pernet O, Beaty SM, Juelich TL, Smith JK, 
            Zhang L, Wang YE, Vigant F, Gao J, Wu P, Lee B, Freiberg AN
  TITLE     Efficient reverse genetics reveals genetic determinants of budding 
            and fusogenic differences between Nipah and Hendra viruses and 
            enables real-time monitoring of viral spread in small animal models 
            of henipavirus infection.
  JOURNAL   J Virol. 2015 Jan 15;89(2):1242-53. doi: 10.1128/JVI.02583-14. Epub 
            2014 Nov 12.
  PUBMED    25392218
REFERENCE   2  (bases 1 to 7385)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/JVI.02583-14"; journalName: "J Virol"; date: "2015-01-15- 
            15"; volume: "89"; issue: "2"; pages: "1242-53"
FEATURES             Location/Qualifiers
     source          1..7385
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          646..1662
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1586..1608
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1670..1689
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4389..4408)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4454..4509
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4508..4527)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4870..4886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4870..4886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4883..4905)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4894..4910
                     /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(4918..4948)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4963..4984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5043..5238
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5089..5224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5151..5170
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5244..5378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5281..5300)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5335..5354
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5446..5463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5617..6205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5697..5716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6376..7236)
                     /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     6999..7018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7237..7341)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(7372..7385,1..366)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"