LOCUS       Exported                6990 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pGloSensor-20F cAMP, complete sequence.
ACCESSION   EU770615
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Stecha PF, McNamara B, Binkowski BF.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAY-2008) Scientific Communications, Promega 
            Corporation, 2800 Woods Hollow Rd, Madison, WI 53711-5399, USA
REFERENCE   2  (bases 1 to 6990)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6990
                     /organism="Expression vector pGloSensor-20F cAMP"
                     /mol_type="other DNA"
                     /db_xref="taxon:549404"
     CDS             complement(5..865)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance protein"
                     /protein_id="ACF75743.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5..865)
                     /gene="ampR"
                     /label=ampR
     misc_feature    970..1123
                     /label=synthetic transcriptional pause site
                     /note="synthetic transcriptional pause site"
     polyA_signal    970..1018
                     /note="synthetic polyadenylation signal"
     misc_feature    1032..1123
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1072..1091)
                     /label=RVprimer3
                     /note="RVprimer3"
     regulatory      1137..1878
                     /regulatory_class="promoter"
                     /note="CMV immediate early enhancer/promoter"
     enhancer        1274..1653
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1654..1857
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1890..1909
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     promoter        1890..1908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1922..4027
                     /codon_start=1
                     /product="GloSensor cAMP"
                     /label=GloSensor cAMP
                     /protein_id="ACF75745.1"
                     /translation="MPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSGYV
                     NNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQH
                     PNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVD
                     EVPKGLTGKLDARKIREILIKAKKGSSGGSGGSGGGMYESFIESLPFLKSLEFSERLKV
                     VDVIGTKVYNDGEQIIAQGDSADSFFIVESGEVKITMKRKGKSEVEENGAVEIARCSRG
                     QYFGELALVTNKPRAASAHAIGTVKCLAMDVQAFERLLGPCMEIMKRNIATYEEQLVAL
                     FGTNMDIVAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVDITYA
                     EYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERE
                     LLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHL
                     PPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQI
                     IPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPT
                     LFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAI
                     LITPEGV"
     regulatory      4060..4281
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late poly(A) signal"
     polyA_signal    4069..4190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4329..4747
                     /regulatory_class="promoter"
                     /note="SV40 early enhancer/promoter"
     promoter        4384..4741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4592..4727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4772..5809
                     /codon_start=1
                     /gene="hygR"
                     /product="hygromycin resistance protein"
                     /label=hygR
                     /protein_id="ACF75744.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     gene            4772..5809
                     /gene="hygR"
                     /label=hygR
     RBS             5788..5796
                     /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)"
     misc_feature    5833..5881
                     /note="synthetic poly(A) signal/transcriptional pause site"
     polyA_signal    5833..5881
                     /note="synthetic polyadenylation signal"
     primer_bind     5948..5967
                     /label=RVprimer4
                     /note="RVprimer4"
     rep_origin      6205..6241
                     /label=ColE1-derived
                     /note="ColE1-derived"
     rep_origin      complement(6208..6795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"