LOCUS       sequence-408464-        8710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pnpr-4 snb-1::pHluorin unc-54 3' UTR C.elegans AVA and other neurons
            expression of snb pHluorin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8710)
  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 8710)
  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: pJH4783
FEATURES             Location/Qualifiers
     source          1..8710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4406..5122
                     /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(4436..4464)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5059..5079
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6150..6172)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6272..6291
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6369..6387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6455..6559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6560..6628,6629..7420)
                     /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(6778..6797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7591..8179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8080..8099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8333..8350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8467..8488
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8503..8508,8509..8526,8527..8533)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8541..8557
                     /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     8546..8568
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8565..8581
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8565..8581
                     /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 acgggagtac ccgggatgga cgctcaagga gatgccggcg cacagggagg aagtcagggc
     3961 ggaccacgac catccaacaa acgtctccag cagacacaag ctcaggttga tgaagtcgtc
     4021 ggaatcatga aggtaagatt ttgaaatgtg caattaaaat aacaaaaatc gcaatttttg
     4081 aaatgatccc atttacaatt tataagcgtg ttaaattaat cttaattccc ctttttgcaa
     4141 taaaatcacg aaaaatagtt ttcaggtgaa cgtggaaaaa gttttggagc gtgatcagaa
     4201 gttgtctcaa ctggacgacc gagctgacgc tctccaggaa ggtgcttcac aatttgagaa
     4261 atctgcggcc accctgaagc gcaagtattg gtggaagaac atcaaaatga tgatcatcat
     4321 gtgcgctatc gtcgtcattc ttattatcat catcgtttta tgggctggag gaaaactggt
     4381 acctatggtg aagggcgagg agctgatgag taaaggagaa gaacttttca ctggagttgt
     4441 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     4501 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     4561 actacctgtt ccatggccaa cacttgtcac tacgttaact tatggtgttc aatgcttttc
     4621 aagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     4681 cgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     4741 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     4801 agatggaaac attcttggac acaaattgga atacaactat aacgatcacc aggtgtacat
     4861 catggcagac aaacaaaaga atggaatcaa agctaacttc aaaattagac acaacattga
     4921 agatggaggc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatgggcc
     4981 cgtcctttta ccagacaacc attacctgtt tacaacttct actctttcga aagatcccaa
     5041 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     5101 catggatgaa ctatacaaat aagctcagca ttcgtagaat tgtaagggat gaactataca
     5161 aataagctca gcattcgtag aattgtaagt ttaaactttc tcatactaac taacgcgctc
     5221 tatttaaatt ttcagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
     5281 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
     5341 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
     5401 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     5461 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     5521 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     5581 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     5641 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     5701 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     5761 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     5821 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     5881 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     5941 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     6001 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     6061 acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt
     6121 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     6181 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     6241 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     6301 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     6361 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6421 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6481 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6541 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6601 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6661 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6721 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6781 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6841 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6901 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6961 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7021 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7081 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7141 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7201 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     7261 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     7321 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     7381 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     7441 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7501 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7561 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7621 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7681 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7741 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7801 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7861 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7921 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7981 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8041 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8101 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8161 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8221 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     8281 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     8341 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     8401 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     8461 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     8521 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     8581 ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta
     8641 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg
     8701 gaaatgaaat
//
