LOCUS       sequence-255884-        7831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C. elegans codon-optimized FlincG3 (GFP-based cGMP sensor) expressed
            in C. elegans ASE neuron pair.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  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 7831)
  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: pSRW1JZ
FEATURES             Location/Qualifiers
     source          1..7831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2774..2791
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2801..2845
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2861..2878
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2897..2911
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(5409..5431)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5531..5550
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5632..5650)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5718..5822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5823..5891,5892..6683)
                     /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(6041..6060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6854..7442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7343..7362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7596..7613
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7730..7751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7766..7771,7772..7789,7790..7796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7804..7820
                     /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     7809..7831
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7828..7831,1..13)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7828..7831,1..13)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaacagctat gaccatgatt acgccaagct tgcatggcag cgcttgactt ctgatgacat
       61 atcaagaatt aaagattttc caaaatgcaa cccatgtttg attcaaggat atgttgttgt
      121 ttccagaaac acaacataat gcaaattcac tcaatacttt ctttctaaag taaatctttg
      181 attaactttc agaattgtaa gaaaaaccaa acaatttcag aattaatcca gaagcaggaa
      241 actgcataaa aaaacaacaa aaaaaaataa tctttctgac aaatgttgac aaaaattatc
      301 gtaaatttac aattttctaa aaatcttcta gatttctttt ccctatgacg gtcgcattaa
      361 agtactgaaa taacataata aaactttaat aataaatttc ttattagaaa attacgattt
      421 tttcattttg acacttcaca gctaactgaa atttttagca cttaattgtc tatgtttttc
      481 caacttcaaa gatcaattta accttcgaaa aacacctgat cacctgaaac tcaatgtttc
      541 aatttcattg ttcaattatt aatgatgtag tacaaattta acaaacacaa aattttccca
      601 tcttcacgtc acttctgctt acataaccta aaatttcagg acgcttttat ttttgaaaga
      661 tccggaaagt gaaatactag ttctgggatt ttgcaagtcc tgcaaaatga aaaaaaatct
      721 atggcttcat cttggttttg aaaaattcca aaaaaaagtg attccatgaa tcagatcagg
      781 aagtactgtt cttttatttt ttaaaataac atatataaat cttcttttta ttttattttc
      841 ttaaaaagtt aatccaaaaa aaagaattgt ttttctccag ggaacggcag taaatcaaat
      901 gacgcagtgg atggatcagt gttgggatgt tcctggaatt gtccacccta taaccaaaac
      961 aaaaagctga ttggaatatt gcttgaaaaa tctgaaacta agatgatttt ctgaaaaacc
     1021 gaagaagttt agaagatcat atatttcccg aataacgcta taatatttat tgaactaact
     1081 ttaaaatatt ctttaggaaa gtataaatcg cgagatataa aattcttaaa tcctagttca
     1141 attgtgataa tgtctaaaaa atgactaata agaaaatatg agaattgttc tatatgcgtc
     1201 attgacattt tcaaaagaag ccttattaga gcagacggat tagatgagta ctcataattt
     1261 aaaaaaacga taactgaaaa tcatacgatc tatcaaagtt tcctaaatgt taaccaaaga
     1321 gtatcgaaaa ttggcaacaa gattcagcac gaaaacttcc tgattttctc cgaacttctg
     1381 atttcatctc acattagctg gaatttaaag ctgaccgaaa aaatagttta acttaaatga
     1441 atattgcgca agtaaatgtt tatttatttt aaagtgagac agcttctaat tattttgttt
     1501 gttttcggtc gatgcaccat gttttgcatt gggttcagtg gatgtattga tgtggaataa
     1561 aagttaatct actgtcagat tgttgactgg gtttttttaa aataccataa gatcctacac
     1621 ttaacgaaac agtgacaggg cacagttttg aaatgtgtgg tgcatatgtg aattgatgat
     1681 tttagtgacc caattgcctt taatgttaat attggttcaa gtgaacgttt gttctcgttc
     1741 aaggtgtctc ataaacatgc ttggaaaatc tcaaaaatga ttttcccaca aagtccataa
     1801 aaattaatta atttgttctt tgactcattt tcactgcaca tttggttcct tctctcacgt
     1861 tctcttactt ttttgtattt tttgaaatct gaaaattgga gtatgagctg cgatacaagt
     1921 cactttgaat ctcgaaatga atccatcaaa tttccttgtt tgatgtcatc aatcttattt
     1981 gctcttcttc aatgtttcgg tgaaaagtca gaaatattgc aattctcaat attctctatg
     2041 gtaactccgt gtttttgttt tgtctggctt ttgactcagg ttaggatcca cctataaaac
     2101 taactttttt accagttaaa catgacgtca atcttccttt ctcatcccca aattcggatt
     2161 agcccccacc aaaaccagca aaaagacggt gaagggcggg aaagagccgt tggagaaagg
     2221 aataccttca acagcttgac tccgcccact tcttcgtccg cttctcctcc cctcaaaaaa
     2281 gcgagtcggg ggtctcattc ggaggctctt ttctaagcca atatcatctt ctttttgctc
     2341 ttttgtagta gtactaatag ataataatat taatcttctt attttcaacc aacttttcac
     2401 ctggtactcc ccctcatcca aattcagaca tgttccacgc cccacgttac cagatgatat
     2461 ttatgaaaag ttgaaaatgt acttcataca acctgacctt cgcagacgta tagctgaatc
     2521 tctatgtttt tctgttcaac gttttttatt tagttattac tcatttttgt tttatatgaa
     2581 aaacaaggta taataggtat aatatttgct tcctgcttta aggattctta tctcataaaa
     2641 tcctgaaatc cattcctgcc ttcttatctt gttttcagtt tttaatcatt attttgaaaa
     2701 caatatgatt attccagaat aagcttgcat gcctgcacta gaggatcccc gggattggcc
     2761 aaaggacatg gcacaccacc accaccacca ctcgtcagga aaggagacag cagcagcaaa
     2821 gtttgagcga cagcacatgg attcaccacc accatctgga cttgttccaa gaggatcagc
     2881 tggatcagga actatcgatg atgatgataa gatggttttg agacaagcat tccgaaaatt
     2941 tacaaagtcc gaacgttcaa aagatttgat taaggaggca atcctggata acgatttcat
     3001 gaagaacctt gaattgtcgc aaattcagga gatcgtggat tgtatgtatc cagtcgaata
     3061 cggaaaagat agttgcatta tcaaggaagg agatgttgga tcacttgtct atgttatgga
     3121 agatggaaag gtggaagtca caaaagaggg agttaagctg tgtactatgg gaccaggaaa
     3181 agtgtttgga gaacttgcaa ttttgtacaa ttgcactaga accgccacag ttaaaactct
     3241 cgttaacgtg aagctgtggg ctatcgatcg acaatgtttc cagactatta tgatgcgtac
     3301 cggattgatc aagcatacag aatatatgga gttccttaaa tctgtgccaa cttttcaatc
     3361 gttgccagaa gagattctct caaaactggc agatgtcctc gaagagactc actatgaaaa
     3421 tggagagtac attatccgtc agggagccag aggagatacc ttctttatca tctcaaaggg
     3481 aaaggtcaac gttacaagag aagattctcc aaacgaagat ccagttttcc ttcgaacatt
     3541 gggaaaggga gattggtttg gagaaaaagc actccaagga gaggatgttc gtaccgccaa
     3601 tgtgattgct gctgaagctg tgacatgcct tgtcattgat agagattcgt ttaagcatct
     3661 tatcggagga ttggatgatg tcagtaacaa ggcttatgaa gatgctgagg caaaagccaa
     3721 gtacgaagct gaggccgctt tctttgcaaa ccttaagaac gtgtacatca aggctgataa
     3781 gcaaaagaat ggaatcaagg caaacttcaa gatcagacat aacatcgaag atggaggagt
     3841 tcagttggca tatcactacc aacagaatac tccaattgga gatggaccag tgcttttgcc
     3901 agataaccat tacctttcta cacaatcgaa attgagtaaa gatccaaacg aaaagcgaga
     3961 tcacatggtc ctcctggagt ttgttacagc agccggaatt actcttggaa tggatgaatt
     4021 gtacaaggga ggaactggag gatcaatggt ttcaaaagga gaagagcttt tcacaggagt
     4081 tgtgccaatt ctcgttgaac tggatggaga tgtgaatgga cataaatttt cagtttctgg
     4141 agaaggagag ggagatgcta cctacggaaa actcacactg aagttcattt gtacaactgg
     4201 aaaattgcca gttccgtggc caactctcgt taccacactg acctatggag ttcaatgctt
     4261 ctccagatac ccagatcaca tgaaacagca cgatttcttt aagtcagcaa tgccagaagg
     4321 atacatccaa gagcgtacaa tcttctttaa ggatgatgga aactacaaga ctagagctga
     4381 agtgaaattc gagggagata ccctcgtcaa tcgaattgaa ctgaaaggaa tcgattttaa
     4441 ggaggatgga aacattcttg gacacaaatt ggaatataac ctgtcggatt tcaacatcat
     4501 cgataccctc ggagtcggag gattcggacg ataagaacat tttcaggagg acccttggct
     4561 agcgtcgacg gtaccatggt attgatatct gagctccgca tcggccgctg tcatcagatc
     4621 gccatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg
     4681 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt
     4741 tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa
     4801 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata
     4861 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat
     4921 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt
     4981 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat
     5041 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt
     5101 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc
     5161 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     5221 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     5281 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     5341 ataactgttt ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac
     5401 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5461 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5521 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5581 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaattaag ggcctcgtga
     5641 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5701 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5761 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5821 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5881 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5941 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6001 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6061 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6121 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6181 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6241 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6301 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6361 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6421 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6481 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6541 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6601 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6661 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6721 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6781 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6841 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6901 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6961 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7021 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7081 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7141 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7201 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7261 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7321 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7381 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7441 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7501 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7561 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7621 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7681 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7741 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7801 gaattgtgag cggataacaa tttcacacag g
//
