LOCUS       sequence-441797-        5471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hlh-34 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 5471)
  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 5471)
  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: pEY99
FEATURES             Location/Qualifiers
     source          1..5471
                     /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             1146..1166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1191..1211
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1233..1253
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3062..3084)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3184..3203
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3281..3299)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3367..3471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3472..3540,3541..4332)
                     /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(3690..3709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4503..5091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4992..5011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5379..5400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5415..5420,5421..5438,5439..5445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5453..5469
                     /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(5458..5471,1..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 tgcctgcagg aagttcatgg
      181 acagttgagt aagtgtattt tgactccaag gcaacgtatg cctgcaccag gagtgtttcg
      241 taaaatttac tgggctacag aaattgttgt atttttctaa tagactcaaa actataccaa
      301 aactgaattt cataatgaga cctcttgaaa atgtctttca aaaaatttca gttgtggttt
      361 aaagtttcga aaactgagaa aaaagaccca cactaatgct aaaataagaa aatcaaaaaa
      421 aaaaaaccga acttcacaaa aaactacgaa agttacgcag gtatgatacc tgcagatgtg
      481 cctgcctacc acataatcga tagttttaac tgcttatttt ttttaggatt atgtgaagcc
      541 gaaggatcat gtcgacattg tgcagacagt agatattcat actatcgatg ccgtatgcag
      601 gatgactgtt ttattggcga aatctgtgat aatggatttt gctgtcccaa cgttttgcca
      661 acattttctc ttcaacgaga acgtgagttt ctatcttgtt ttattttttg cctacaatta
      721 actatcaata cttttactaa cataatctaa cacactgcac tatttctcag caactagtcc
      781 taaaccaaaa agaccattaa atatcacatc ggatccccca gctgatgagc catgtccaga
      841 tgaatctaga tggtcaaaaa ggtgctcaag agatgaggat tgtactcatc aagaagaact
      901 ttgcatcgag ggaaaatgtt gtaaaagtgc gtttattttt taaatttgaa accaaagtgt
      961 ttatcttctt tgaaattagc gtaataaaaa aagaaataaa atgagaaccc gggattggcc
     1021 aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga
     1081 cccttggagg gtaccggttt tcccagtcac gacgcgtggt accgagctca gaaaaaatga
     1141 ccgctccaaa gaagaaacgc aaagtaccga gctcagaaaa aatgacggca cctaaaaaga
     1201 aacgcaaagt accggtcgcc gagaagacag ccccaaaaaa gaagcgtaaa gttccacgcg
     1261 tcatgggagt aagtttaaac agttcggtac taactaacca tacatattta aattttcagc
     1321 gcgctgtctc caagggagag gagctcatca aggagaacat gcacaccaag ctctacatgg
     1381 agggaaccgt caacaaccac cacttcaagt gcacccacga aggggagggc aagccctacg
     1441 agggcaccca gaccaacagg attaaggtgg tggagggagg ccccctgccg ttcgcattcg
     1501 acatcctggc cacctgcttt atgtacggga gcaagacctt catcaaccac acccaaggaa
     1561 tcccagactt cttcaagcaa tccttcgtaa gtttaaacat gattttacta actaactaat
     1621 ctgatttaaa ttttcagcct gagggcttca catgggagag agtcaccaca tacgaagacg
     1681 ggggcgtgct taccgttacc caggacacca gcctccagga cggctgcttg atctacaacg
     1741 tcaagctcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     1801 gctgggaggc cagtaccgag accctgtacc ccgctgacgg cggcctggaa ggcagatgcg
     1861 acatggccct gaagctcgtg ggcgggggcc acctgcactg caacctgaag accacataca
     1921 gatccaagaa acccgctaag aacctcaaga tgccaggagt aagtttaaac atatatatac
     1981 taactaaccc tgattattta aattttcagg tctactttgt ggaccgcaga ctggaaagaa
     2041 tcaaggaggc cgacaatgag acctacgtcg agcagcacga ggtggctgtg gccagatact
     2101 gcgacctccc tagcaaactg gggcacaaac ttaatggcat ggacgagctg tacaagtaag
     2161 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc
     2221 agagtaagtt taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc
     2281 gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct
     2341 ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt
     2401 ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta
     2461 attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc
     2521 ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt
     2581 ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt
     2641 atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca
     2701 tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt
     2761 aattttcctg cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg
     2821 cgtttttctt gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag
     2881 gtttgggttt gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt
     2941 gtctatgggg tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt
     3001 ttcctactag tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3061 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3121 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3181 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     3241 gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt
     3301 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3481 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3541 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3601 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3661 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3721 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3781 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3841 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3901 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3961 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4021 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4081 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4141 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4201 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4261 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4321 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5161 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5221 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg atgaagcgga agagcgccca
     5281 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5341 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5401 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5461 ggataacaat t
//
