LOCUS       sequence-292696-        7597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This construct contains cGMP biosensor driven by nlp-1 promoter for 
            expression in PHB sensory neuron in pSMdelta backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7597)
  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 7597)
  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: MKV937/nlp-1p::FlincG3
FEATURES             Location/Qualifiers
     source          1..7597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2532..2549
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2559..2603
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2619..2636
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2655..2669
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(5167..5189)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5289..5308
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5390..5408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5476..5580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5581..5649,5650..6441)
                     /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(5799..5818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7354..7371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7488..7509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7524..7529,7530..7547,7548..7554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7562..7578
                     /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     7567..7589
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7586..7597,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7586..7597,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcgcc actactgccg agttgtgctc
       61 actgcaattt tctaactttt tactttctac tttttacttt tcactttttt actcgagctc
      121 tagttcttct ttcatcaaaa tattcgaatg tccgggaggg atctatcagt cacgaggtgg
      181 actttgacac ctttaataat gtggttttcg ttcttcctga tggagtgctt gctggttttg
      241 acagcaactt tttggaacct atttcatttt gttgcgcgtt tagtttaagg atgacgattt
      301 ctatctgaat ttattacaat tatctgatct agtggcacaa actccgggag atcaaacaca
      361 cacaatagag aagattaact catcagtttt tgtgtgcccc ccgtcccttc aaagctttat
      421 tccagctttg ttttatccaa cattattcac aagtgccagc ctctaatatt cttcatttct
      481 gattctcatc tctattagcc aattttgttt gactcacacg tcatttgatc cattagccaa
      541 gttgccacct caaaggacat ttcattggat tagccataat gaataatctc ccaaaaaact
      601 ttaatgctcc tcctcacatc atctcagtct ctgtgccatg attgttttct tctatcagtt
      661 gtctattgct gctagcctgg agaaggtgtg tttgatttgt tctataatat gggaacggaa
      721 atcaattaca ttactctcct cctgtttgta ttatatccaa tcattttcta tattactgct
      781 acaattttgg gtcactgact cattgctgaa tgtgatagct taaacacgtg acacctttct
      841 tggtgaatcc accacaattt ttcttcaaat ctatgttgag taatcggatt aatctattac
      901 tcggttttga atgctatgtt ttacattttt tactatggtg caaaaattac caaatttcat
      961 agaaaaagtg ttttgaaaaa tttttattga cagtcggaaa tactcagttt tttttccatt
     1021 ttttgaaaaa aaaaactata gccatgttgc atattcaaat ccctacaatg ttttaatatt
     1081 attacttgaa acatacgtag atcataaaat tcaaaagaaa ttttttttga tttttatttg
     1141 gcgacttcca aaaaatggca aatatctttt ttttaacttt tcgaaaaaat aatttgaaat
     1201 ttttaaattt tgaaaaattg aaaactataa tactttgtac taacctagcc atgaaaaatt
     1261 gaatgttgca cattttgact tgctactttc ctccattcaa tattcatctg aattcaaaat
     1321 agtaaagcag gcaattttgg aattttagac tggtaaatga tgccgctctc aagaattgtc
     1381 tactcaaagg atccaggaag aaaaataaga aataccctta attattaacc ggatttggtc
     1441 tttgggaaga tcacactttc tttgtgacaa cctatgtaaa aattccaatt agtttcatct
     1501 tcttcactca tattcaagta ttcccaaagc gcgaaataag cgaatgatca attgtttttt
     1561 gttactcatg ttattctaag tattactcat ttattcatga tcattaagag tcataattgt
     1621 aatagagcac ttttgaatga aaatcgtgat ttgagatgtc aaaaatacac tttttcacct
     1681 ccctcgataa tttctataat ttttcactac acgtcatttc ccacttccgc tcatatcatt
     1741 agaaaaccaa gcggagacaa tgacttggcg atattagcat atcccttagt cacaaattcc
     1801 aagttccatg atttcacaca tatttataaa ttctgatctc gaattcaatg agtcccgaat
     1861 aatccggcat ttgaaacatt gtgctccacc ctttgtttac atattcattt ctttctagct
     1921 ccattttttg tggcctgtgc ggcagaattg tgcgtaccca gagcattttg gataaggtgc
     1981 aacaaatcaa acatcaacct ttcaaaaccc cttaatcctg tttttgatta caacacaagt
     2041 ctcgaaagtg ttgttagctc aacctttcgc ccaatctcat cccataaaac atataacttt
     2101 tggtatacat ttgattggct ttggcaattc ccggaagaaa acgctttcag acttttgatt
     2161 cttaccgtat tacgttcaat gtttttcttt ccggaagtgt ttaaattttt tcctctgtcc
     2221 cattttcgtc atgcacgaac accaagaaac cgtagttcag ttgttcagtt ttgtcttgca
     2281 tttcccaacg gccctatatt ttcaaaaaaa aaggttattt cttagaaacc actcaaaagt
     2341 ccagcccatt ttttctcctc taaaattgca aaaaattgtc aatttctcaa cactcataag
     2401 ccccgccttc cacgatttcg catggtttta tgttttccct aaattgtact tttcatatat
     2461 ttcctgacat ttcagacatc aacttgaggc aacgtctaga ggatccccgg gattggccaa
     2521 aggacatggc acaccaccac caccaccact cgtcaggaaa ggagacagca gcagcaaagt
     2581 ttgagcgaca gcacatggat tcaccaccac catctggact tgttccaaga ggatcagctg
     2641 gatcaggaac tatcgatgat gatgataaga tggttttgag acaagcattc cgaaaattta
     2701 caaagtccga acgttcaaaa gatttgatta aggaggcaat cctggataac gatttcatga
     2761 agaaccttga attgtcgcaa attcaggaga tcgtggattg tatgtatcca gtcgaatacg
     2821 gaaaagatag ttgcattatc aaggaaggag atgttggatc acttgtctat gttatggaag
     2881 atggaaaggt ggaagtcaca aaagagggag ttaagctgtg tactatggga ccaggaaaag
     2941 tgtttggaga acttgcaatt ttgtacaatt gcactagaac cgccacagtt aaaactctcg
     3001 ttaacgtgaa gctgtgggct atcgatcgac aatgtttcca gactattatg atgcgtaccg
     3061 gattgatcaa gcatacagaa tatatggagt tccttaaatc tgtgccaact tttcaatcgt
     3121 tgccagaaga gattctctca aaactggcag atgtcctcga agagactcac tatgaaaatg
     3181 gagagtacat tatccgtcag ggagccagag gagatacctt ctttatcatc tcaaagggaa
     3241 aggtcaacgt tacaagagaa gattctccaa acgaagatcc agttttcctt cgaacattgg
     3301 gaaagggaga ttggtttgga gaaaaagcac tccaaggaga ggatgttcgt accgccaatg
     3361 tgattgctgc tgaagctgtg acatgccttg tcattgatag agattcgttt aagcatctta
     3421 tcggaggatt ggatgatgtc agtaacaagg cttatgaaga tgctgaggca aaagccaagt
     3481 acgaagctga ggccgctttc tttgcaaacc ttaagaacgt gtacatcaag gctgataagc
     3541 aaaagaatgg aatcaaggca aacttcaaga tcagacataa catcgaagat ggaggagttc
     3601 agttggcata tcactaccaa cagaatactc caattggaga tggaccagtg cttttgccag
     3661 ataaccatta cctttctaca caatcgaaat tgagtaaaga tccaaacgaa aagcgagatc
     3721 acatggtcct cctggagttt gttacagcag ccggaattac tcttggaatg gatgaattgt
     3781 acaagggagg aactggagga tcaatggttt caaaaggaga agagcttttc acaggagttg
     3841 tgccaattct cgttgaactg gatggagatg tgaatggaca taaattttca gtttctggag
     3901 aaggagaggg agatgctacc tacggaaaac tcacactgaa gttcatttgt acaactggaa
     3961 aattgccagt tccgtggcca actctcgtta ccacactgac ctatggagtt caatgcttct
     4021 ccagataccc agatcacatg aaacagcacg atttctttaa gtcagcaatg ccagaaggat
     4081 acatccaaga gcgtacaatc ttctttaagg atgatggaaa ctacaagact agagctgaag
     4141 tgaaattcga gggagatacc ctcgtcaatc gaattgaact gaaaggaatc gattttaagg
     4201 aggatggaaa cattcttgga cacaaattgg aatataacct gtcggatttc aacatcatcg
     4261 ataccctcgg agtcggagga ttcggacgat aagaacattt tcaggaggac ccttggctag
     4321 cgtcgacggt accatggtat tgatatctga gctccgcatc ggccgctgtc atcagatcgc
     4381 catctcgcgc ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct
     4441 ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc
     4501 tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata
     4561 gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat
     4621 tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa
     4681 atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt
     4741 cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca
     4801 tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat
     4861 gaaattaatt ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga
     4921 ggacggcgtt tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg
     4981 aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg
     5041 agtagtgtct atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat
     5101 aactgtttct actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5161 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5221 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5281 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5341 gcacagatgc gtaaggagaa aataccgcat caggcggcct taattaaggg cctcgtgata
     5401 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5581 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5641 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5701 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5761 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5821 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5881 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5941 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6001 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6061 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6121 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6181 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6241 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6301 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6361 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6421 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7321 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7381 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7441 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7501 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7561 attgtgagcg gataacaatt tcacacagga aacagct
//
