LOCUS       Exported                6308 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  NanoLuc CMV-Hygro luciferase vector pFN31A, complete sequence.
ACCESSION   KF793052
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6308)
  AUTHORS   McNamara B, Kopish K, Robers M.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-OCT-2013) Scientific Communications, Promega 
            Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA
REFERENCE   2  (bases 1 to 6308)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6308
                     /organism="NanoLuc CMV-Hygro luciferase vector pFN31A"
                     /mol_type="other DNA"
                     /db_xref="taxon:1454401"
     regulatory      1..742
                     /regulatory_class="promoter"
                     /note="CMV immediate-early enhancer/promoter; EcoICRI site 
                     removed"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          857..989
                     /note="chimeric"
     regulatory      1033..1052
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1607
                     /codon_start=1
                     /gene="NLuc"
                     /product="NanoLuc luciferase"
                     /label=NLuc
                     /protein_id="AHH41346.1"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILAGSSGAIAME"
     gene            1065..1607
                     /gene="NLuc"
                     /label=NLuc
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    1578..1589
                     /gene="NLuc"
                     /label=linker
                     /note="linker"
     misc_feature    1590..1597
                     /gene="NLuc"
                     /note="used in Flexi vector cloning; SgfI site"
     CDS             1621..1956
                     /codon_start=1
                     /product="barnase"
                     /label=barnase
                     /protein_id="AHH41344.1"
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     misc_feature    1958..1965
                     /label=PmeI site
                     /note="PmeI site"
     regulatory      2117..2338
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyA signal"
     polyA_signal    2126..2247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2437..2849
                     /regulatory_class="promoter"
                     /note="SV40 early enhancer/promoter"
     promoter        2492..2849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2700..2835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2880..3917
                     /codon_start=1
                     /gene="hygR"
                     /product="hygromycin resistance protein"
                     /label=hygR
                     /protein_id="AHH41345.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     gene            2880..3917
                     /gene="hygR"
                     /label=hygR
     RBS             3896..3904
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     regulatory      3941..3989
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyA signal"
     polyA_signal    3941..3989
                     /note="synthetic polyadenylation signal"
     promoter        4145..4249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4250..5110
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /function="resistance to ampicillin"
                     /label=ampR
                     /note="ampicillin-resistance protein"
                     /protein_id="AHH41343.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4250..5110
                     /gene="ampR"
                     /label=ampR
     rep_origin      5265..5301
                     /label=ColE1-derived plasmid replication origin
                     /note="ColE1-derived plasmid replication origin"
     rep_origin      complement(5268..5856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5972..6255
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"