LOCUS       sequence-362023-        8020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Exogenous EP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8020)
  AUTHORS   Watabe T, Yamahira S, Takakura K, Thumkeo D, Narumiya S, Matsuda M, 
            Terai K
  TITLE     Calcium transients trigger switch-like discharge of prostaglandin 
            E(2) in an extracellular signal-regulated kinase-dependent manner.
  JOURNAL   eLife. 2024 Jan 26;12:RP86727. doi: 10.7554/eLife.86727.
  PUBMED    38276879
REFERENCE   2  (bases 1 to 8020)
  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: "eLife."; 
            date: "2024-01-26"; volume: "12:RP86727. doi"; pages: " 
            10.7554/eLife.86727"
COMMENT     Sequence Label: pPBbsr2-PTGER2
FEATURES             Location/Qualifiers
     source          1..8020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2692..2711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    4083..4632
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4227..4244)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4454..4473
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(5104..5121)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5110..5334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5381..5397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5405..5421
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5429..5435,5436..5453,5454..5459))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5474..5495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5729..5791
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5832..5852)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5832..5850)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5871..5887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5895..5911
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5919..5925,5926..5943,5944..5949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6102..6119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6273..6861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6353..6372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7032..7823,7824..7892))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7655..7674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7893..7997)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg gsccscsccv gggggggggg gggggggggg ggggsggggs ggggcgaggc
     1561 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2701 tggttattgt gctgtctcat cattttggca aagaattggg atcagatttc ctggaattct
     2761 gcagatatcc agcacagtgg cggccgcgcc accatgaaga cgatcatcgc cctgagctac
     2821 atcttctgcc tggtattcgc cgactacaag gacgatgatg acgccagcat cgatatggga
     2881 aacgcttcta atgattccca aagcgaagac tgtgagacca ggcagtggct cccccccggt
     2941 gaatctccag cgatttcctc cgtcatgttc tctgccggtg ttctcggtaa cttgatcgcg
     3001 cttgctctcc tcgctaggcg ctggagaggc gacgtggggt gcagcgcagg gagacgctca
     3061 tcactgagtc tgtttcacgt gctggtcact gaattggtct tcaccgatct cttgggcaca
     3121 tgtctgatct cccccgtggt gctggctagc tatgcccgca atcagacatt ggtcgccttg
     3181 gccccagaga gccgcgcttg tacctatttc gctttcgcca tgacattttt ttctctcgca
     3241 acaatgctca tgctgtttgc catggctttg gagcggtacc tgagtatcgg ccacccatat
     3301 ttctaccagc ggcgggtcag caggtccggg ggactggcag tgctcccagt gatttacgcc
     3361 gtgtcactgt tgttttgttc actgccgctg ctggactatg gtcagtatgt gcaatactgt
     3421 cccggaacat ggtgttttat tcgacatggg aggacagcct acctccaact ctacgctaca
     3481 ctgttgctgc tcctcatcgt gtccgtcctg gcctgcaatt tctctgtaat tctcaacctc
     3541 attcgcatgc atcggagaag ccgccggagc aggtgtggcc caagcctggg atctggacgc
     3601 ggtgggccag gtgcaaggcg ccgcggagaa agggttagca tggcagaaga aacggaccac
     3661 ctgattctcc tggcgataat gaccatcact tttgccgtgt gcagccttcc ttttacaatc
     3721 ttcgcataca tgaacgagac cagtagtaga aaagaaaagt gggacctgca ggctttgcga
     3781 tttctctcta tcaactctat cattgacccc tgggtttttg ccattctgcg ccctccagtc
     3841 ctgaggctga tgcgctccgt cttgtgttgt agaataagcc tgaggaccca ggacgccacc
     3901 cagacaagtt gcagtacaca gagcgatgct tccaagcaag ctgatctcat cgataccggt
     3961 taaaggtctc gagggatcga gggcggccgc tctagagtcg acgggccgcg gtaacaattg
     4021 ttaactaact taagctagca acggtttccc tctagcggga tcaattccgc cccccccccc
     4081 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     4141 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     4201 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     4261 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     4321 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     4381 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     4441 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4501 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     4561 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4621 aacacgataa taccatggtc atgaaaacat ttaacatttc tcaacaagat ctagaattag
     4681 tagaagtagc gacagagaag attacaatgc tttatgagga taataaacat catgtgggag
     4741 cggcaattcg tacgaaaaca ggagaaatca tttcggcagt acatattgaa gcgtatatag
     4801 gacgagtaac tgtttgtgca gaagccattg cgattggtag tgcagtttcg aatsgacaaa
     4861 aggattttga cacgattgta gctgtkagac ancccttatt ctgacgaagt agatagaagt
     4921 attcgagtgg taagtccttg tggtatgtgt agggagttga tttcagacta tgcaccagat
     4981 tgttttgtgt taatagaaat gaatggcaag ttagtcaaaa ctacgattga agaactcatt
     5041 ccactcaaat atacccgaaa ttaaaagtcg agatgcatcg atgatctaga gctcgctgat
     5101 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5161 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5221 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5281 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5341 aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5401 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5461 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5521 tccagtcggg aaacctgtcg tgccagcgga tcttaaaagt tttgttactt tatagaagaa
     5581 attttgagtt tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat
     5641 ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat
     5701 aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa
     5761 cgtacgtcac aatatgatta tctttctagg gttaatctag tatacgcgtt taattaatcc
     5821 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     5881 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5941 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6001 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6061 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6241 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7021 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7081 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7141 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7201 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7261 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7321 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7381 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7441 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7501 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7561 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7621 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7681 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7741 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7801 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7861 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7921 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7981 caaatagggg ttccgcgcac atttccccga aaagtgccac
//
