LOCUS       sequence-355751-        9701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prgef-1 NLF-1::GFP unc-54 3' UTR C.elegans pan-neural expression of 
            NLF-1::GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9701)
  AUTHORS   Xie L, Gao S, Alcaire SM, Aoyagi K, Wang Y, Griffin JK, Stagljar I, 
            Nagamatsu S, Zhen M
  TITLE     NLF-1 delivers a sodium leak channel to regulate neuronal 
            excitability and modulate rhythmic locomotion.
  JOURNAL   Neuron. 2013 Mar 20;77(6):1069-82. doi: 
            10.1016/j.neuron.2013.01.018.
  PUBMED    23522043
REFERENCE   2  (bases 1 to 9701)
  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"; doi: 
            "10.1016/j.neuron.2013.01"; journalName: "Neuron"; date: 
            "2013-03-20- 20"; volume: "77"; issue: "6"; pages: "1069-82"
COMMENT     Sequence Label: pJH2198
FEATURES             Location/Qualifiers
     source          1..9701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4335..4363)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5111..5131
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6828..6845)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6828..6844)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6837..6859)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7057..7512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7144..7163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7354..7375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7529..7548)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7648..7670
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7708..7726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7794..7898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7899..7967,7968..8759)
                     /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(8117..8136)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8930..9518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9419..9438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9672..9689
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttcgaatggc
      241 catgggacgt cgacgcttgc atgcctgcag cgtttccgat acccccttat atcagcacac
      301 attcagtcat cataccacaa aatgttgtga tagtgatagg caaaactacc gtatttcctc
      361 tattaatctt gcatgcaaga ctaattttcg attaacccgt aggggtgcaa gactaataga
      421 gactgcaaga ctattagagg ctgaaatact aattttcgta tgctcaataa ttttggaaat
      481 tggcctattt tttgtagaaa cttgataccg tttaaacaag gaaaaataca cactttttaa
      541 tatttcatca ataatttgaa cgattttgtg attttaagtt caatttgccc aaaaaaagac
      601 aatttttctg acgttaccgc aaagtaatgc tggtcaggca aaaagagcgg tgcaaaaata
      661 taagagactg caatactaat agaggaaata cggtaattga attttagtga aacttgcgac
      721 agttttcctc atttttttgt tattccggca tacgagtgtg gcatacgagt gaggtcatct
      781 tttgtttctt ccgtttcttc atcgcttttg aaaaaaaatg ttgaagaaaa cttcgtagcc
      841 gatagccgga taatgtttgg cacggcgttc caacatatat cattggaaat tttttaagat
      901 ttttgcgaaa aatcacattc ttcacgatga gaacacgtta ttgaaggaat ataaatcaag
      961 aataacatat agttatattt ctctattact ttaacgttaa atatgagcaa atttgagcat
     1021 tttgattgcg atgaaaagca gaatcgagtc aactgaaatc cgttcaaaaa cctaagctcg
     1081 gttcgcccga aagtcgattt ttccaatagc cgaacagcac tgcttcttta ttttccaggt
     1141 atttttgggg tgtaagaatt atatcttctt tgtttgggta tttacgtttc cgaattccct
     1201 ttcttgatct gtcgctattt ttcagaaacc agaacagttt atcctttttc atgagaacct
     1261 gaatagctca aaaacccgtt taatttcttt tcatcatctc atttgaaact ttcctagtct
     1321 tcttaatgat ttccaggctc ctcctacatt tgtatctcaa gatgcataca ctttatttcc
     1381 gctccaattc gaatcatcta cttctctttt ttcttcaact ttctactttt ttccattctt
     1441 ttctttcaaa tgttcgacgt cttcaagtga ctgctcttca ttctcctcat ttttctcgaa
     1501 tttcattctt gtctcttttt tgctcaaaaa tggaaaatga aagtgacgtg agatttggac
     1561 ggcgggacac gggggcagta gaagcagcaa aaaggagaga aagaggacac aaataagaag
     1621 aacgaattca aaaataagcg gagaggagct atttccgtca attctacctc cccaatcttc
     1681 atcaattcgg ctcaattgaa tgacgtcaca gggagataaa ccgtttggat gagcgccgtc
     1741 catcactgac gccatcccgt ttgggacaag aaaagagaaa aaagagcaca aagtttttgg
     1801 tgacggatct tgtcaatcat atgaaagttg ttctgattga ttgtcagttt ttttcctact
     1861 tttttggatt ctatccactt ctgaactttt gacaagtttc aaactttctg aatcattttc
     1921 tatgcatttt cctggaattc tttttatgta aaatatgaaa tagaatgttt ttgaattcaa
     1981 gttctgcttt tttcttcttt tctgttctgt tcgggcttgg gtatgctttt tttaaaaatt
     2041 attttgcaca tcgaccaata agtgcgcaac ttataaaatt aatttatttt tgttaatttt
     2101 tgaaatactt gtattgcttt aagtgatctg acctcgccct gagctttcca cgtagttatc
     2161 aaatacaaat cctccaaggg taacgtacct atattactga tctttataat aactttatca
     2221 cctgtccagt tccagaggaa ttctgttaag cttataggca cagaaggagt catttctgct
     2281 ggtttttaat gatccaaatc tttatttcaa gtaaaaaact gaacacttgc gaataaaact
     2341 atcagattaa ccattcacca aaaatgtgtt tgaatctaaa actttctcag tattccaaat
     2401 atagaaataa ataaccacga cattgctaaa atctgtctga attgtgtact ccttaccgtg
     2461 aagtaataat ggatataatg aatcgtttga aatgaatgat cagcacattt ttggtgaaag
     2521 atcacaaata aggaataagc gtcggaaaat aaaacgatta tttcggatca aaaatttgtt
     2581 gaagatcata tacactcgag accaagatta ttctagacaa ttttcaaatt ggcttctttg
     2641 tttgcaaatc ttcataataa tctgtgaagt ttggaaattt gaatttttaa tcttttttca
     2701 tagattatag tttttatttc tttgcaaaac tatattaaaa accgatgcat tgttttaggg
     2761 aaattaatga gccttttgtt caacactaaa aacaataaaa ttaaaatttt ggcttcatca
     2821 tttgaccttt tttaagttcg aaaacttttc tcgtatttct gaaccgccaa ttttttcaca
     2881 catctctaga cttttggtgc ccgttccaga aagttaagta attgctattc taagaaagtt
     2941 ctcaacattg ttttttagtt ctgattgaat tctgatgttc caggaatata ttttaaattt
     3001 aatattcttt gcactatttc tatactaact aaataataaa tagtctaagt atgttcaatg
     3061 agaacaaaaa agctctatct atatttttct atcctatttc atttatatcc ttttcatttt
     3121 gaactcacca gcttctcttc ttcttcttgt ccatatactt cttaaaagtc tcgaacttct
     3181 ctctctcact ctccatcaat ttctttatgg ctaccgcttg cgctctgccg cttccgaaga
     3241 aagaaggcgt tggcagagct ctcaattcta ttttttctcg ccgtaggcta tcgatttctc
     3301 agcttctctt ctctcttcct cactccacca ccaccacttc gggcttcttc ttttcttcat
     3361 tttcgtcttc ttcttcatcc tcttcttttt ttttcagttc cccctcactc ctccccttat
     3421 atgcgcgtgc gagaggggtg caaaagcagc gcgcatccga gaattgaaac gagaaaacgg
     3481 agacgcagca gcagttcgtc ctcagaaaaa tagccagtaa aaagagaaaa agatagagag
     3541 agacctatcg cattttattt tcaaattgca attcctgcaa ttcatgtgtg cgtgtgccca
     3601 ttttcaattc ttcccgttat ttttcagacc attaccaacg ttctttctag atcttgagac
     3661 aatcttcctt ctgctcaatc gttcgtcgtg aagacggcat cgacgacgac gacgatcagc
     3721 agtaaaggat ccttatgcgg cctcgcacgc cttacacaaa agaagcgcta cgatacgagc
     3781 gtcgtcatca aaagcctcca cccactcatc cgcctgctcc tacagctgct cctcgtccag
     3841 acactacaga agtattcggc cggcgagaag caaattcgtc gccgatcacc gattttatcc
     3901 agtcgttttg tgcgagcatt caagacgaaa gctgcagttc aactgacgag ccaaactttg
     3961 aattcaaaaa tgatattact gaacgattat ggttaaagcc ttttgatatt gtgccttcgc
     4021 ctcaaatttt atccgattcg aagaaaacta gttctccaag aaggtgctgg gccaagaatt
     4081 ttcgccgata tttaaccgat ggtattctta ttattcaatt atttacttta ttcaattgcg
     4141 cttcttcatc tcatttatcc aacacaattc ctccttgtaa aaattttggg ttttttgcgc
     4201 agcatagcga caatgtaact gatcgttgca cagtttggtc tgatgacgtc ccgcggatta
     4261 atataattat gagtcgacaa ggaattcaag taccggtaga aaaaatgagt aaaggagaag
     4321 aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca
     4381 aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat
     4441 ttatttgcac tactggaaaa ctacctgttc catgggtaag tttaaacata tatatactaa
     4501 ctaaccctga ttatttaaat tttcagccaa cacttgtcac tactctcact tatggtgttc
     4561 aatgcttctc gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
     4621 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
     4681 gtaagtttaa acagttcggt actaactaac catacatatt taaattttca ggtgctgaag
     4741 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     4801 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca
     4861 tcatggcaga caaacaaaag aatggaatca aagttgtaag tttaaacatg attttactaa
     4921 ctaactaatc tgatttaaat tttcagaact tcaaaattag acacaacatt gaagatggaa
     4981 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     5041 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     5101 gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggacg
     5161 aactatacaa aagcccacaa gcttcagggc ccgagctcga cgtcattcct gaaatttgct
     5221 caactggcac acaatcacgt attagaatat taaagcatgt gttccttttt ccagagttac
     5281 aaccgttatc attttttgaa atattcgccg tcggcttgag aaacactgac acgtggcagt
     5341 tgaatgccgg cgagaagggt aatgattgca tttcgggaag tataaatcgg tgtacatcgt
     5401 gtttccggag gatctcaggc actcttcgta agcttaatga agcctatcgg tctttcgaca
     5461 caactctttc tcgtttcgac tgccttccag cggtggacac tgcctccgca acaagaccgt
     5521 tctcgccgaa cgcaacttgt gataattgta aaatatggta tcgtagatgg ttgttggttc
     5581 aatcgcttca aatctggcaa cggcctccgt gtattaactg gtgctactat actcaattgg
     5641 catgcccaca tctcgcacct gcaaaattat gggactacgc aggtcatccg tcgtttcagt
     5701 gtagagatat ggacattgac agttggcgcc acgaatgtga ctgcatccat ccgtgtgatg
     5761 ttcgtggaat tgtagctcca ggtgttgcaa tgggcaacgc tcctttagtt cagcacgact
     5821 ttttcgcggc acagattcac tgtgaaacgc ggaaaaagga gtgccacaag cgtttcaaac
     5881 agattcggaa atcaaaacgg agtgacacca gcagtaacat tgttttttta tcaagcgcat
     5941 caccaaaacg cccaaaattc ctgattcttt cattgttgtt tcttcgattt atgttgttgt
     6001 tatagggggt accatggtat tgatatctga gctccgcatc ggccgctgtc atcagatcgc
     6061 catctcgcgc ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct
     6121 ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc
     6181 tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata
     6241 gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat
     6301 tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa
     6361 atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt
     6421 cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca
     6481 tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat
     6541 gaaattaatt ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga
     6601 ggacggcgtt tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg
     6661 aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg
     6721 agtagtgtct atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat
     6781 aactgtttct actagtggcc acgtgggccg tgcaccttaa gcttggcact ggccgtcgtt
     6841 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     6901 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6961 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     7021 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     7081 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     7141 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     7201 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7261 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7321 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7381 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     7441 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7501 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7561 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     7621 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     7681 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     7741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     7801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7921 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7981 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     8041 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     8101 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     8161 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     8221 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8281 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     8341 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     8401 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     8461 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     8521 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     8581 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     8641 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     8701 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     8761 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     8821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9541 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9601 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9661 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga g
//
