LOCUS       sequence-441594-        7636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tol-1(3k) fluorescent neural reporter driving nuclear mNeptune2.5 
            expression (refer to NeuroPAL paper for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7636)
  AUTHORS   Yemini E, Lin A, Nejatbakhsh A, Varol E, Sun R, Mena GE, Samuel ADT,
            Paninski L, Venkatachalam V, Hobert O
  TITLE     NeuroPAL: A Multicolor Atlas for Whole-Brain Neuronal Identification
            in C. elegans.
  JOURNAL   Cell. 2021 Jan 7;184(1):272-288.e11. doi: 
            10.1016/j.cell.2020.12.012. Epub 2020 Dec 29.
  PUBMED    33378642
REFERENCE   2  (bases 1 to 7636)
  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.cell.2020.12.012"; journalName: "Cell"; date: "2021-01-7-
            7"; volume: "184"; issue: "1"; pages: "272-288.e11"
COMMENT     Sequence Label: pEY138
FEATURES             Location/Qualifiers
     source          1..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..22
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6..22
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(1370..1384)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             3311..3331
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3356..3376
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3398..3418
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5227..5249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5349..5368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5446..5464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5532..5636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5637..5705,5706..6497)
                     /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(5855..5874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6668..7256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7157..7176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7544..7565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7580..7585,7586..7603,7604..7610)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7618..7634
                     /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     join(7623..7636,1..9)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta
       61 ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac
      121 gatggatacg ctaacaactt ggaaatgaaa taagcttgca tgcctgcaga caagtacggc
      181 aaatcggcaa ttcgccgatt tgccggattg ccggaaatct tgattttcgg caaaccggca
      241 aacatcagcg tactatttta ctattcaaaa taaatgtagg aacattcata ggatgcgtac
      301 aattttgccg ataaaattta aattctgaag cttcaaaaaa aatgtgcaaa accacaattt
      361 gccgaaaatt ctagccgatt tcaattccgg caattttttg ccgaaaaaaa ttgccgccca
      421 cccctgattt atattcagtc tgttaccgat tctagtgagg ggtttccagc ctttgcatga
      481 ataggtgtct gctacattaa ttgcgcaatc cacattgata cagagcaacc tcaattgttg
      541 ttttttctgt ggcccgttct gctcgttaca cctataaaaa ggtggtcaaa caagtcgtaa
      601 aatttgggtc atgagatggt ccctgggtta attggagaag tgccgtcatt gagagagaga
      661 gagccaatta cgtatgaggt ctgctctgct ctcgggaaga ctctataacc cttcgttttt
      721 ggcgggagat atgagatatt ttgctgtaat tatcgcactt gttttgggta ttctcttttt
      781 gtatgattta cctaaaattt ttgagcagtt gtgattatcc tattttgttt cgagaaccta
      841 gaaatgtttc ctatcatagt aaccgtttaa ctctgtgagt atagttttac tttaagttgc
      901 tccgtttttt atttgactag cacagctttt ttacattgct ccggttttgg ccaaaggttt
      961 tctgtttttt ttttgcagtt ctgacttatt ctggcctgct ttacccagct aagcctaggc
     1021 ccggttacag tccataagac aacatttttg aactcaataa ttgccgaatt gccaaaatgc
     1081 caaattttcg attgctgccc gtctgagtaa gtctgaaaca gtttgtgtga aattgagccg
     1141 gaaactactc ggagtgatgt ggccacttcc ctatttagtt tctcggttga ttttcacact
     1201 tcccgccgag agcccgaagc atctgccagc tgaacggttt ccgtcgtttt tcatgtctat
     1261 cgctaaaatg tgcgcgttac acacacataa acatcacttc tctcataaag tgatggataa
     1321 ttcgagtgga ttgcatctga caccacactt ccgataatcg cttcgtcgtc ttgtcatcat
     1381 cgtcgttttc ttcgtcttgt tgtcgtccgg tccccacaca cacatcaaaa cggctacttc
     1441 cgacgtgtcg gtggtttgtg tgtgtgtcaa acggcaccac cacaagtggt tgccctcgcc
     1501 cgtgtgctaa ttcgcatttt gtcttgttaa tcacgcaata gtgataacaa tcgcttccgg
     1561 accattgcta atataatata ttggctaccg cgggcatgtc ttggcaataa ccttgatgtc
     1621 aagttttatc gttaaaatgt gaagctgtat gattgcgaaa aatttgaagt tttcccattc
     1681 aaattgtagg catatttgaa aaatttctga agatttatat ctgtcgtaag tggaacggtt
     1741 tatttggttt cttgggtctc gtcgcgaaaa gctttcacta tggtttgaag cacgactggg
     1801 cgacaatcga agtcgaagtt cggcaattcg ataattcggc aattatcgag ttcaagattt
     1861 tcgaaagaaa aactcaaaaa ataattttga aatctattta ttagttctgg acacgcggag
     1921 tcagaaagtc ccatttcgct ttgatctacg aaaaatgcgg gagttgagac gcagacatct
     1981 caactgattt cgcatggtta agagtgtgct gacgtcacaa ttgttctgga aaaaaattcc
     2041 cgcatttttt gtagatcaaa ccgtaatggg atagcctgga cgtagaactc gccaaatcgt
     2101 agatatatgc ataatgaaaa ctctagagtt aactctgcca ccctcataaa gtgagttaga
     2161 aacatttctg ctctgaggaa ctatagaacc ccctttcaag tcgatcgtca gcaaattcta
     2221 gaaagatctg acagtggccg attttttaat gtttctaggc cacgtaaaaa agctgatgtc
     2281 ttgttacttt tcaattcgaa aatccctttt tttaattttt cggcagcacc cggtccgagc
     2341 caacatcttc ctaaacactt caaaaccccg cccccttctg ccctcccggg tgtctccgtg
     2401 ttgttcaggg tgttgtccac cccctagaca cccaactgac cattcacccg tctcctcctc
     2461 cccctctaca acaaccacca ccacctgatc cattccatta taatgatatt ccgcatactt
     2521 gtcatacaga gaatacccgg tttgctgctg ccaaacggtt ttcatttatt ccatttctca
     2581 ctccttgatt gttgtcaatt cttcgattcg atgcggcaaa actagcgcca gtggggaaat
     2641 tgctttaata gtaaacaatt tttttttaaa tttcctatat tttttgtata agattttctt
     2701 ttttaattgc aaatcccatg ggaaagtctc aaattgcata aaattccaat ttgaattccc
     2761 tccaagattt caatgttcga tggagcgcac ttgaattatt ataattttta tcaatgaatt
     2821 tttttatcat ctctgactga tttttcacga ttttttgtgt agttttagag gaaatttact
     2881 gaaaaatcca agttaaatgt aaatttccga tttttataca atttttggac tactttcccc
     2941 gccgccaccg ctaaaaatcc caatttcctg caagttttca cctgctctga ctcatttccg
     3001 cgagcgcgcg cgctcgtact ttttatcttt tactatttat ttatctcctc caaaaaaaaa
     3061 tcccgttaat tttttttccc ttacaatttc tcataattta ctcatttagt ttggcttaaa
     3121 aaatgctaaa ttaaattttt ttttcatttc cagatcacat gacacaacga aacccgggat
     3181 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
     3241 gaggaccctt ggagggtacc ggttttccca gtcacgacgc gtggtaccga gctcagaaaa
     3301 aatgaccgct ccaaagaaga aacgcaaagt accgagctca gaaaaaatga cggcacctaa
     3361 aaagaaacgc aaagtaccgg tcgccgagaa gacagcccca aaaaagaagc gtaaagttcc
     3421 acgcgtcatg ggagtaagtt taaacagttc ggtactaact aaccatacat atttaaattt
     3481 tcagcgcgct gtctccaagg gagaggagct catcaaggag aacatgcaca ccaagctcta
     3541 catggaggga accgtcaaca accaccactt caagtgcacc cacgaagggg agggcaagcc
     3601 ctacgagggc acccagacca acaggattaa ggtggtggag ggaggccccc tgccgttcgc
     3661 attcgacatc ctggccacct gctttatgta cgggagcaag accttcatca accacaccca
     3721 aggaatccca gacttcttca agcaatcctt cgtaagttta aacatgattt tactaactaa
     3781 ctaatctgat ttaaattttc agcctgaggg cttcacatgg gagagagtca ccacatacga
     3841 agacgggggc gtgcttaccg ttacccagga caccagcctc caggacggct gcttgatcta
     3901 caacgtcaag ctcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac
     3961 actcggctgg gaggccagta ccgagaccct gtaccccgct gacggcggcc tggaaggcag
     4021 atgcgacatg gccctgaagc tcgtgggcgg gggccacctg cactgcaacc tgaagaccac
     4081 atacagatcc aagaaacccg ctaagaacct caagatgcca ggagtaagtt taaacatata
     4141 tatactaact aaccctgatt atttaaattt tcaggtctac tttgtggacc gcagactgga
     4201 aagaatcaag gaggccgaca atgagaccta cgtcgagcag cacgaggtgg ctgtggccag
     4261 atactgcgac ctccctagca aactggggca caaacttaat ggcatggacg agctgtacaa
     4321 gtaagcgccg gtcgctacca ttaccaactt gtctggtgtc aaaaataata ggggccgctg
     4381 tcatcagagt aagtttaaac tgagttctac taactaacga gtaatattta aattttcagc
     4441 atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc ctacatgctc
     4501 tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct cttaatttct
     4561 ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag
     4621 aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt aataataatt
     4681 ctatcccaaa atctacacaa tgttctgtgt acacttctta tgttttttac ttctgataaa
     4741 tttttttgaa acatcataga aaaaaccgca cacaaaatac cttatcatat gttacgtttc
     4801 agtttatgac cgcaattttt atttcttcgc acgtctgggc ctctcatgac gtcaaatcat
     4861 gctcatcgtg aaaaagtttt ggagtatttt tggaattttt caatcaagtg aaagtttatg
     4921 aaattaattt tcctgctttt gctttttggg gtttccccta ttgtttgtca agatttcgag
     4981 gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag aaagatcgga
     5041 agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt tgaaagtgga
     5101 gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat accaaacata
     5161 actgtttcct actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5221 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5281 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5341 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5401 gcacagatgc gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc
     5461 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5521 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5581 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5641 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5701 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5761 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5821 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5881 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5941 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6001 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6061 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6181 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6481 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6781 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7321 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7381 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgatgaa gcggaagagc
     7441 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     7501 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     7561 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     7621 tgagcggata acaatt
//
