LOCUS       Exported                4793 bp ds-DNA    circular SYN 26-7-2013
DEFINITION  Promoterless vector for measuring the activity of promoter and 
            enhancer sequences with a secreted NanoLuc(R) luciferase assay.
ACCESSION   JQ513380
VERSION     .
KEYWORDS    pNL2.3[secNluc Hygro]
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  AUTHORS   Promega
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4793
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     misc_feature    1..70
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             187..699
                     /codon_start=1
                     /gene="
                     "
                     /product="NanoLuc(R) luciferase"
                     /note="Nluc"
                     /note="human codon-optimized"
                     /protein_id="
                     "
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    748..869
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        1063..1420
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      1271..1406
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             1451..2488
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     polyA_signal    2512..2560
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(2887..3475)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3675..4535)
                     /codon_start=1
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4640..4688
                     /note="synthetic polyadenylation signal"
     misc_feature    4702..4793
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"