LOCUS       sequence-412783-        8558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pnpr-4 ins-22::pHluorin unc-54 3' UTR C.elegans AVA and other 
            neurons expression of ins-22 pHluorin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8558)
  AUTHORS   Meng J, Ahamed T, Yu B, Hung W, Ei Mouridi S, Wang Z, Zhang Y, Wen 
            Q, Boulin T, Gao S, Zhen M
  TITLE     A tonically active master neuron modulates mutually exclusive motor 
            states at two timescales.
  JOURNAL   Sci Adv. 2024 Apr 12;10(15):eadk0002. doi: 10.1126/sciadv.adk0002. 
            Epub 2024 Apr 10.
  PUBMED    38598630
REFERENCE   2  (bases 1 to 8558)
  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: "Sci Adv."; 
            date: "2024-04-12"; volume: "10(15):eadk0002. doi"; pages: " 
            10.1126/sciadv.adk0002. Epub 2024 Apr 10"
COMMENT     Sequence Label: pJH4687
FEATURES             Location/Qualifiers
     source          1..8558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4254..4970
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4284..4312)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4907..4927
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5998..6020)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6120..6139
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6217..6235)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6303..6407
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6408..6476,6477..7268)
                     /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(6626..6645)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7439..8027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7928..7947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8181..8198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8315..8336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8351..8356,8357..8374,8375..8381)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8389..8405
                     /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     8394..8416
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8413..8429
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8413..8429
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcatgc ggccgcatca tgacatcagt attctgctat tcagctaaga
       61 ttcttgttgt cgtcatttga tgtatccgtt cgtcggtgca ggacttccac atggcgcagt
      121 cgaaaccatg ttaatctctg aagtcaaatg gtctaagaat tcacagtata gacggactgc
      181 aggtgtaaac ttatcctata tatcacccaa gacgtattac cttgtcagac tgcatttaca
      241 caagtaatcc cacttattat atgatgtcac gcgttataaa acggaagctt acttattgtg
      301 gaacaggtgg aatactcaaa aacattgaaa attcttacaa tttctaagta aaaaccttat
      361 gaatgttttg tacgtattat tattcccatt cttccattta gttgtcagtt tttcattgct
      421 cgggaagatt tgcataaata tcgccattga ccagtctact ccccagtttt ttgatcatga
      481 tttttcacaa caatgatcaa tcaaataaac atcatatcat cttatcccga ttaagcgccc
      541 tcgcaatttt attgcctcat cttcttaata atagaaacca tatgctcatt tttggcttag
      601 tttttgctca gttcaatttt ccttattcca tcgtaagcgt taagtctaga aggctttttg
      661 ttattcactt tttttctttg ctttttactt ttctctttct tcacgagaca aatattggta
      721 ttattgctca tctctctact aataaaactc aacaatttcg ttgaaacttg gtctctagag
      781 tttgatgagt tgtttgtgtt atttttgatc taaattttca attttgtgga agtaaggctt
      841 gcaagataga caaagacaac aagaaaataa acgaagggat tgaggaagaa taaactgaaa
      901 tgatcatggt taagaaaata ggaagaaaat agaagaagag cggccggctc actaggttaa
      961 attatttttg tgcttgttgt ggggagcgtt gagtaagacc tagagcaagg aacgcggtta
     1021 gcaatccctg ccgtttccaa atctccagag actatgcgaa attcagaaga cacgggacat
     1081 caagacagat aggtacggag taagaccttc aaattgtttc ttctcttctt gattactctc
     1141 ttgctcctct ctcgtccatt gtgctctttc ccacacactg agcaaaagca tcgtcattgt
     1201 ggaaagtgag cttgatgtgg ttggctcaaa catctgacca gctagacttg tcacttgtaa
     1261 gatatcacat cgcacttccc aattaatctt aaaatccgct tgactcattt atttttgaat
     1321 tcgtttttgt ttgaaattgt aaacttcgga atagttccag accgattcag aactagtatg
     1381 atcacctatg aaacgtgatt tctgaaatat tcttactttt ttctagacaa ctcaagatct
     1441 cgatcggcca aaattgtatt atcctcgtaa tcatcataac catttccgat attctgatca
     1501 caacggcttt ctttcgaatc tagtacgtct cacgtacaac tgcccacaaa cgatttccat
     1561 gaaactgata aataattaaa gactttggac ttaccaagag ttcaaacagg ttttatttcg
     1621 ttttctattt ttgaaaaaaa aatcattcca gttggacgac aaaaacagtt gtagaacttt
     1681 aataataata tttattcttg acacaaaaaa acttatcgta tttgtaaggg gaaaatagac
     1741 ccataattaa caacgtttct attagttatg cattttactg tactcatgat attactccaa
     1801 taacaaataa tttatctgaa atagcacttt tctccttctc cctgtgctcc aatacagtcc
     1861 agacacatat taattttcaa caccacatca cgcacatctt ccataggaaa ttaaaaaaaa
     1921 aatacattag acttcctatt catatcaaat atttcacatc ataatatttt tgtttgctca
     1981 tttttcaaca cttttctgat tcattttatt tttgtacgtt acgataccta catcagttat
     2041 ttttcacctg ccgtcactta ttttagatat tccttaattt tgactacgta ttactcattg
     2101 ttaacatttc tgatatttgt atactcaaag ccatttggaa agaatcagaa cagtgatttc
     2161 agttttgatg ctggccaact agttgaataa cgtgatgagg accattgagt aattagagaa
     2221 aacatgttta cgaatatagt cggctaaatt gagaaaaata cctaaataac tggcaataag
     2281 ctcagaactg aaaaacaaaa aattgctaac catcccgtga atttttgatg tgtgcctgtc
     2341 tatgccgtca tgttcatctc atgtgctttg ttttcaatcc ctggtctctt gtaatttttc
     2401 ttttaaaaag aagtctgcta ttcgtgacaa attaaaattg aaaattcata tatctttaga
     2461 gtatagccgt tttgagtcag caacaaacaa ttaggtgcca cttccgggac aggcacctct
     2521 gaaataaaag gaataagaaa atagtgagat attcccccct cactgatcaa acttcttttt
     2581 ttccctttac tacgtttacc tatattttta ttctgttagt ttggtcgaaa attggtggag
     2641 gatgtctaga atttgcagga tacaaattga gactaaagaa aattgaattt cttcaagtct
     2701 tctcgatttc ttgatttcta gtaaagcttc ctttgtaact actgtaatag ttagtgagaa
     2761 taattaaatt ttggcgcctg ccagtcattt ctagtgaaca aatataacat caatactttg
     2821 aaatctaatt ttctaagtag ctcgggtaat ttttttgttc acatcactcc ttacaacaca
     2881 actttcttca aaaatttcac gtctatattt agtttcaatt tttcaaaggt tgaaggcgtc
     2941 tagacttcga tgaataccaa aattaaagca atcatagttt tttaggtaaa gtttatgaat
     3001 ctgcaaagtc atactataaa actgatctta gaaatttttc ttgcagattt tatcttaata
     3061 tttcaaaaaa catcgttatg ttcccatttt gatttttggt gtaatggtcg tgtttctacg
     3121 aacaaaaacg gagacacccc caaaaatttt tgagatgctg tttttgaaaa taataaaggc
     3181 agctgtcagc tttattcatt tttctgaaaa acaattcact gtcattttga gctaatagct
     3241 gcaaaaccaa caaacgtaaa aactcagaaa gcattcaaaa atcgtaaaac ttgagttttt
     3301 ctcaagaaat acgattaaga ttactgtaga caaatttata gtaaattgaa ttaaagtgta
     3361 atttgataaa tgcatgactt tctcagattt attcattttc cacatataaa ctctaattga
     3421 atactaggaa ctgtgtatga gtatatcatt tgttgaaatg ctacattttt ctgttagtct
     3481 agatttttta acgagtttgt gtcttggtaa ttgtcgtgtg tattcattgt tcacagcttc
     3541 taataggaaa gggttatttt caaaaaacgt tctgtacttc gctggtagga gtactgtagt
     3601 tttactattt tttttatata acagaactat ttgtttctgc tttgaaagca taattcgaat
     3661 aattacgtag gattcaattt ttcttgatac tcattttctt ttttgtctaa ccatttcatt
     3721 ctgacacata aattaccaat aacagattga ttgtctgtac tttatgttat tttcctttca
     3781 atatttttcc tctctcttat ttctaaaagc ggaaatgaaa aacgactctg agaattaatt
     3841 ttcttcattt cttctgtagg cgtaactgca ctttctggga tttctttttg ctgtcataat
     3901 acggagtacc cgggatgcac actacaacta ttctcatatg ctttttcatc cttcttgttc
     3961 aagtctccac aatggatgct cacactgaca aatacgtcag aactctgtgt ggaaaaactg
     4021 caatcagaaa tattgccaac ctttgcccgc caaagccaga aatgaagggt atctgttcta
     4081 ccggaggtga gataaactta aactcaaaag ccttgtttgc aaataattat ttttctaaat
     4141 tttcagagta tccaagcatc accgaatact gttccatggg attttcagac tctcagatca
     4201 agtttatgtg ctgtgataac caatgggtac ctatggtgaa gggcgaggag ctgatgagta
     4261 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     4321 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     4381 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
     4441 cgttaactta tggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact
     4501 ttttcaagag tgccatgccc gaaggttacg tacaggaaag aactatattt ttcaaagatg
     4561 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
     4621 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
     4681 acaactataa cgatcaccag gtgtacatca tggcagacaa acaaaagaat ggaatcaaag
     4741 ctaacttcaa aattagacac aacattgaag atggaggcgt tcaactagca gaccattatc
     4801 aacaaaatac tccaattggc gatgggcccg tccttttacc agacaaccat tacctgttta
     4861 caacttctac tctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     4921 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa gctcagcatt
     4981 cgtagaattg taagggatga actatacaaa taagctcagc attcgtagaa ttgtaagttt
     5041 aaactttctc atactaacta acgcgctcta tttaaatttt cagaattcca actgagcgcc
     5101 ggtcgctacc attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag
     5161 taagtttaaa ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc
     5221 ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct
     5281 gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt
     5341 agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc
     5401 gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa
     5461 aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga
     5521 aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga
     5581 ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt
     5641 gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt
     5701 ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt
     5761 tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt
     5821 gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct
     5881 atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc
     5941 tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6001 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6061 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6181 cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat
     6241 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6301 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6361 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6421 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6481 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6541 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6601 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6661 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6721 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6781 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6841 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6901 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6961 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7021 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7081 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7141 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7201 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7261 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7321 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7381 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7441 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7501 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7561 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7621 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7681 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7741 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7801 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7861 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7921 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7981 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8041 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8101 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8161 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8221 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8281 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8341 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     8401 aacaatttca cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat
     8461 gatcttacta actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac
     8521 tcacaacgat ggatacgcta acaacttgga aatgaaat
//
