LOCUS       sequence-471815-        7104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express -42F cGMP in biosensor assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7104)
  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            "Unpublished"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 -42F cGMP Vector
FEATURES             Location/Qualifiers
     source          1..7104
                     /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    4183..4304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4248..4267
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4498..4855
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4498..4689
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4521..4541)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4706..4841
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4768..4787
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4886..5923
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5902..5910
                     /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    5947..5995
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6151..6168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6322..6909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6402..6421)
                     /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 agggaggttc aggcggttcc ggaggttctg gcggatcagg
     2521 cggttcgggg ggctcgagcg gagaattagt 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 tttggcattt tgtggtattg ggagctccgg aggttcaggc ggttccggag gttctggcgg
     3061 atcaggcggt tcggggggag ccaaaaacat taagaagggc ccagcgccat tctacccact
     3121 cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc
     3181 cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt
     3241 cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg
     3301 gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt
     3361 catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag
     3421 catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct
     3481 caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga
     3541 ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa
     3601 cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa
     3661 cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt
     3721 ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat
     3781 cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat
     3841 ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt
     3901 gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa
     3961 gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc
     4021 gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg
     4081 ccagggctac ggcctgacag aaacaaccag cgccattctg atcactccag aaggggttta
     4141 aactctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     4201 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4261 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4321 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     4381 tctacaaatg tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat
     4441 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg
     4501 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4561 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4681 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4741 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4801 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4861 cgattcttct gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga
     4921 aaaatttctc atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga
     4981 agagagccga gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag
     5041 ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact
     5101 acccatcccc gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag
     5161 tagacgcgcc caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt
     5221 acagcctgtc gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg
     5281 cttcggccca ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg
     5341 cgccattgct gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc
     5401 cagcgtagct caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg
     5461 ccacctcgtc catgccgact tcggcagcaa caacgtcctg accgacaacg gccgcatcac
     5521 cgccgtaatc gactggtccg aagctatgtt cggggacagt cagtacgagg tggccaacat
     5581 cttcttctgg cggccctggc tggcttgcat ggagcagcag actcgctact tcgagcgccg
     5641 gcatcccgag ctggccggca gccctcgtct gcgagcctac atgctgcgca tcggcctgga
     5701 tcagctctac cagagcctcg tggacggcaa cttcgacgat gctgcctggg ctcaaggccg
     5761 ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt cgcacacaaa tcgctcgccg
     5821 gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg gccgacagcg gcaaccgccg
     5881 gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt taagatctag aaccggtcat
     5941 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttcga
     6001 actagatgct gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     6061 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     6121 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6481 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6721 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6841 aaccaccgct ggtagcggtg gttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     7081 ggccgcaaat gctaaaccac tgca
//
