LOCUS       sequence-332388-        7813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of bPGC in BWMs of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7813)
  AUTHORS   Woldemariam S, Nagpal J, Hill T, Li J, Schneider MW, Shankar R, 
            Futey M, Varshney A, Ali N, Mitchell J, Andersen K, Barsi-Rhyne B, 
            Tran A, Costa WS, Krzyzanowski MC, Yu YV, Brueggemann C, Hamilton 
            OS, Ferkey DM, VanHoven M, Sengupta P, Gottschalk A, L'Etoile N
  TITLE     Using a Robust and Sensitive GFP-Based cGMP Sensor for Real Time 
            Imaging in Intact Caenorhabditis elegans.
  JOURNAL   Genetics. 2019 Jul 22. pii: genetics.119.302392. doi: 
            10.1534/genetics.119.302392.
  PUBMED    31331946
REFERENCE   2  (bases 1 to 7813)
  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: "Genetics. 
            2019 Jul 22. pii: genetics.119.302392. doi: 
            10.1534/genetics.119.302392."
COMMENT     Sequence Label: pMS04 [myo-3p::bPGC::SL2::mCherry]
FEATURES             Location/Qualifiers
     source          1..7813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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)."
     primer_bind     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3959..4669
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4104..4122)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4372..4391
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5585..5607)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5707..5726
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5808..5826)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5894..5998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5999..6067,6068..6859)
                     /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     complement(6217..6236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7030..7618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7519..7538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7772..7789
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcggc tataataagt
      241 tcttgaataa aataattttc ccgacaaaac atgagtattt ctttcgaaaa taaaagtgca
      301 ggctaattag agattattct gtaattaact gcataatttg tcacgtgcca tagttttaca
      361 ttccactacg tcatagttct taaaatacta atctcctgaa aatagaagta ggtgaagaaa
      421 gtttaattat cagttctaaa atgacaattg atctttggaa tatgttctga aactaccgat
      481 cattgaacag atgctatttg aatgatatag aattgtatat ttgcaatttc tgaaacgcgt
      541 tcttaaaggc acacagatta attcaaaagg gtctggccgc aaaaaggttt atggtggccg
      601 attttgagtt ttgtgtgtga ttgctttttc acaatcagtg ttttcaggat tatgtgatga
      661 actagatctt caagtttcgt tacatttcat atgttttcgg aactcacgaa gtacatattg
      721 ggtattgtgc tcaaaaaatt cagcaatcag cttcgctccg ctgactttag aacccaaaaa
      781 aatagtatgg ccaaactgac tgtgttacga tcatttcaat ttttcaatac atatttaaga
      841 tttctaagag taagaaggtc aaaaactgtt ctggaataca tatatatttt tcaggttaca
      901 attagtcaaa aagtgcactg aaatatacgt tttaatttca cgaataaccc aattagttca
      961 atgtattttt ggtcaaccaa cgttaaagtt tggcttccaa ccaattatca tttctgatca
     1021 accacaatgt tttttcttta tctgcaagtt aattttttat ttttatccag atgtttggca
     1081 tatttttcaa ttcttcacta gcgcccactt cttgcacttc cggcgccctg aatctaatgc
     1141 atctgttgca agaattgaaa gaccaatcaa cacattgttt tcttcacgag atactgaaga
     1201 aaatgaataa aaacagagaa aaagagccat gtgattagtg acaactgttg ctaacagata
     1261 ccatagcttg gacttggtac gtgatggcaa cgtatgggtc aacaaaaatg attgcagagg
     1321 gggtgcaaaa cagtcaagtc gagaaaatat gaaaaacaga aaacaaagaa cagaaaaatg
     1381 ggtttgagag tcagtataat ttataaaaga aaaattgtac atagaaatta accatttttg
     1441 tagaagaagt tatttttcaa gcatcgttaa aaattattca aagcacctta tttcatattt
     1501 aattttaaac atggttaaat gaacaacacg gtgcgcaatc aggaaaactt gaaatctgaa
     1561 actgttgttg tgatcttctt cgcaactgtt cagatagcac tagtgtaatg ttaagagtgc
     1621 gcgaatataa tggaatataa tggatcacac ctcctgccat caggtaaacg tctctgttat
     1681 cacatatttc caactattaa atttttacct tttacagttt tacatttttt tgaaaaaagt
     1741 aactttttgt cttcaaaatc cctgacgaaa atatcaaata ttttaatcga gactgcagag
     1801 gaaccgattg atgatttgga aaatccagct ttacctgtgt aagaactgaa aagtttcata
     1861 accctagggt attcccagtt acattcccca ctggctaaca atagcaccca gtttttcatc
     1921 acctttcttc aaatttctcg gcgatttgtt aaaaacaaaa tttgtgtccc ttctctgata
     1981 tctctatgtc tctaaacaca agttcatcgg aaaacgaagg agggtaggtg ttggttgggc
     2041 tcccgaagtg aaaatagaag agcaagaata gaatattaga gagagagtgc agagagggcg
     2101 ggatagctcc cgggattccg ttttcttctt ctttatcttc aacgatgatg tgtgtgcgtg
     2161 ttgtatagat tctgttgctc ccccacaact cgctccgaag gctcaataca attcaattga
     2221 tattggagga gagcctaccg gagtgggagg ataagaagaa acataagaag aagaagaaga
     2281 agaagcatgc ttctggtttt tgatgctatg aaaacggcac aaaaagatga ttgaggtccc
     2341 ttttcaatac cttctctcat ctttcaaatc ccattgaaac ctaaaacttc tcaccacgct
     2401 ttaccattgt tctccaaaaa cttatagcaa tgtctataac ttttttatct ctgaaaagca
     2461 gtgttccatt tttctttttc ctattttatt tcaattgttt ctcacatttc gtttggattc
     2521 tttgcttgtc aaccagcttc ttcttccact tttaccgtct aattttcagg gcagggagcc
     2581 atcaaaccca cgaccactag atccatctag aggatccaag cttgccacca tgatgaagcg
     2641 gctggtgtac atcagcaaga tcagcggcca cctgagcctg gaagagatcc agcggatcgg
     2701 caaggtgtcc atcaagaaca accagcggga caacatcacc ggcgtgctgc tgtacctgca
     2761 gggcctgttc ttccagatcc tggaaggcga gaacgagaag gtggacaagc tgtacaagaa
     2821 aatcctggtg gacgaccggc acaccaacat cctgtgcctg aaaaccgagt acgacatcac
     2881 cgaccggatg ttccccaact gggccatgaa aaccatcaac ctgaacgaga acagcgagct
     2941 gatgatccag cccatcaaga gcctgctgca gaccatcacc cagagccacc gggtgctgga
     3001 aaagtacatg cccgccagag tgatctacct gatcaaccag ggcatcaacc ccctgaccgt
     3061 ggagccccag ctggtcgaga agatcatctt cttcagcgac atcctggcct tcagcaccct
     3121 gaccgagaag ctgcccgtga acgaggtggt catcctggtc aaccggtact tcagcatctg
     3181 cacccggatc atcagcgcct acggcggcga agtgaccgag ttcatcggcg actgcgtgat
     3241 ggccagcttc accaaagagc agggcgacgc cgccatccgg accagcctgg acatcatctc
     3301 cgagctgaag cagctgcggc accacgtgga ggccaccaac cccctgcacc tgctgtacac
     3361 cggcatcggc ctgagctacg gccacgtgat cgagggcaac atgggcagca gcctgaagat
     3421 gaaacacggc ctgctggggg acgccgtgaa cgtggccgcc aggctggaag ccctgacaag
     3481 acagctgccc tacgccctgg cctttacagc cggcgtgaag aagtgctgcc aggcccagtg
     3541 gaccttcatc aacctgggag cccaccaggt caagggcaag caggaagcca tcgaggtcta
     3601 caccgtgaat gaggcccaga agtactacga caccctgcag atcacccagc tgatccggca
     3661 gaccctggaa aacgacaagt aactagcgtc gacggtaccg ctgtctcatc ctactttcac
     3721 ctagttaact gcttgtctta aaatctatgc ttctctttag tatctaaaat tttcctagaa
     3781 gcttacaagt atataaatgg tctcttctca ataaaggttg tatatttatt catcttattg
     3841 aatctgccat ttcctcgttt ttgcgagttt atataccttc caattttctt tctattgtat
     3901 tttcaacttc taattttaat tcagggaaac tgctgtaccg gtagaaaaaa tgctcgagat
     3961 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     4021 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     4081 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     4141 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     4201 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     4261 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     4321 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     4381 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     4441 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     4501 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     4561 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     4621 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag aattccaact
     4681 gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc
     4741 atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat
     4801 ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt
     4861 tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt
     4921 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     4981 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     5041 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt
     5101 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     5161 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     5221 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     5281 attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga
     5341 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     5401 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     5461 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     5521 tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5581 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     5641 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5701 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5761 acagatgcgt aaggagaaaa taccgcatca ggcggcctta attaagggcc tcgtgatacg
     5821 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6181 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6301 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6541 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6601 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6721 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6781 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6841 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6901 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6961 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7021 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7081 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7141 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7201 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7261 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7321 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7381 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7441 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7501 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7561 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7621 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7681 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7741 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7801 gcgcccaata cgc
//
