LOCUS       sequence-471819-        7092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -40F cGMP in biosensor assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7092)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7092)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGloSensor -40F cGMP Vector
FEATURES             Location/Qualifiers
     source          1..7092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(5..796,797..865))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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     1072..1091
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     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"
     primer_bind     1808..1828
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1854..1878
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1890..1909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1890..1908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4171..4292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4182..4201)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4236..4255
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4486..4843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4486..4677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4509..4529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4694..4829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4756..4775
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4874..5911
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5890..5898
                     /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)"
     polyA_signal    5935..5983
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6139..6156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6310..6897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6390..6409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggttacca gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt
       61 ccatagtggc ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag
      121 gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa
      181 tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca
      241 tccagtctat gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc
      301 gaagagttgt ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt
      361 cgttcaactc tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa
      421 atgcagtcag ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt
      481 cgctcatggt aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct
      541 tttccgtgac cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa
      601 gctgctcttg cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag
      661 tgctcatcat cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga
      721 gatccagttc gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca
      781 ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg
      841 cgacacgaaa atgttggatg ctcatactcg tcctttttca atattattga agcatttatc
      901 agggttacta gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg
      961 acaggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     1021 tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa
     1081 taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctggcctaa ctggcctcaa
     1141 tattggccat tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg
     1201 ccattgcata cgttgtatct atatcataat atgtacattt atattggctc atgtccaata
     1261 tgaccgccat gttggcattg attattgact agttattaat agtaatcaat tacggggtca
     1321 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1381 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1441 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1501 ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt
     1561 aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag
     1621 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat
     1681 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1741 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     1801 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     1861 ttagtgaacc gtcagatcac tagaagcttt aatacgactc actatagggc tagcgatcgc
     1921 catgcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga
     1981 cactggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat
     2041 catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg
     2101 gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg
     2161 gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat
     2221 cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc
     2281 cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga
     2341 gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt
     2401 gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga
     2461 gattctcatt aaggccaaga agggctcgag cggaggttca ggcggttccg gaggaggttc
     2521 tggcggatca ggcggttcgg gggaattagt gaaggatatt tctagtcatt tggatgtcac
     2581 agccttatgt cacaaaattt tcttgcatat ccatggactg atatctgctg accgctattc
     2641 cctgttcctt gtctgtgaag acagctccaa tgacaagttt cttatcagcc gcctctttga
     2701 tgttgctgaa ggttcaacac tggaagaagt ttcaaataac tgtatccgct tagaatggaa
     2761 caaaggcatt gtgggacatg tggcagcgct tggtgagccc ttgaacatca aagatgcata
     2821 tgaggatcct cggttcaatg cagaagttga ccaaattaca ggctacaaga cacaaagcat
     2881 tctttgtatg ccaattaaga atcataggga agaggttgtt ggtgtagccc aggccatcaa
     2941 caagaaatca ggaaacggtg ggacatttac tgaaaaagat gaaaaggact ttgctgctta
     3001 tttggcaggg tccggtggat ccggtggcag cggagggacg tcaggtggat ctggagggag
     3061 ctccggtgcc aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac
     3121 cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc
     3181 ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt
     3241 tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg
     3301 cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc
     3361 tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag
     3421 ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa
     3481 gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact accagggctt
     3541 ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt
     3601 cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag
     3661 taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca
     3721 tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt
     3781 gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg
     3841 ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa
     3901 gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat
     3961 cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa
     4021 ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg
     4081 cctgacagaa acaaccagcg ccattctgat cactccagaa ggggtttaaa ctctagagtc
     4141 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     4201 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4261 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4321 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4381 gtaaaatcga taaggatccg tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc
     4441 atggcctgaa ataacctctg aaagaggaac ttggttagct accttctgag gcggaaagaa
     4501 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4681 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4741 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4801 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga
     4861 cactagcgcc accatgaaga agcccgaact caccgctacc agcgttgaaa aatttctcat
     4921 cgagaagttc gacagtgtga gcgacctgat gcagttgtcg gagggcgaag agagccgagc
     4981 cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg
     5041 cttctacaaa gaccgctacg tgtaccgcca cttcgccagc gctgcactac ccatccccga
     5101 agtgttggac atcggcgagt tcagcgagag cctgacatac tgcatcagta gacgcgccca
     5161 aggcgttact ctccaagacc tccccgaaac agagctgcct gctgtgttac agcctgtcgc
     5221 cgaagctatg gatgctattg ccgccgccga cctcagtcaa accagcggct tcggcccatt
     5281 cgggccccaa ggcatcggcc agtacacaac ctggcgggat ttcatttgcg ccattgctga
     5341 tccccatgtc taccactggc agaccgtgat ggacgacacc gtgtccgcca gcgtagctca
     5401 agccctggac gaactgatgc tgtgggccga agactgtccc gaggtgcgcc acctcgtcca
     5461 tgccgacttc ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga
     5521 ctggtccgaa gctatgttcg gggacagtca gtacgaggtg gccaacatct tcttctggcg
     5581 gccctggctg gcttgcatgg agcagcagac tcgctacttc gagcgccggc atcccgagct
     5641 ggccggcagc cctcgtctgc gagcctacat gctgcgcatc ggcctggatc agctctacca
     5701 gagcctcgtg gacggcaact tcgacgatgc tgcctgggct caaggccgct gcgatgccat
     5761 cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt
     5821 atggaccgac ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc ccagtacacg
     5881 accgcgcgct aaggaggtag gtcgagttta agatctagaa ccggtcatgg ccgcaataaa
     5941 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt
     6001 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     6061 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     6121 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6841 tagcggtggt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
     7081 taaaccactg ca
//
