LOCUS       sequence-375956-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lim-6 fluorescent neural reporter driving nuclear mNeptune2.5 and 
            CyOFP1 expression (refer to NeuroPAL paper for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  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 6314)
  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: pEY49
FEATURES             Location/Qualifiers
     source          1..6314
                     /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             599..619
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             644..664
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             686..706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1610..1663
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1673..1693
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1715..1735
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3905..3927)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4027..4046
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4124..4142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4210..4314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4315..4383,4384..5175)
                     /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(4533..4552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5346..5934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5835..5854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6088..6105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6222..6243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6258..6263,6264..6281,6282..6288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6296..6312
                     /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(6301..6314,1..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 tgcctgcagt tagccagttg
      181 cataaatatt aaagtcaaat gttcaaagga aataatattc tgacccgtgc taatgcaata
      241 tttggtcagt gccaatgaaa atctccattt tctaatgaat tttacgattt gcaaacaaca
      301 aacattccag acagatgcgc gaaatgagat ggctgcctgc taccgtagtg ctttgttgat
      361 ttaacacaca tttcgcttca agcgttttgg tgtttaagaa attgcgattt gaatttggaa
      421 aaaaggttca aatcaaagag aattttagca agaagcttag cccgggattg gccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 agggtaccgg ttttcccagt cacgacgcgt ggtaccgagc tcagaaaaaa tgaccgctcc
      601 aaagaagaaa cgcaaagtac cgagctcaga aaaaatgacg gcacctaaaa agaaacgcaa
      661 agtaccggtc gccgagaaga cagccccaaa aaagaagcgt aaagttccac gcgtcatggg
      721 agtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc agcgcgctgt
      781 ctccaaggga gaggagctca tcaaggagaa catgcacacc aagctctaca tggagggaac
      841 cgtcaacaac caccacttca agtgcaccca cgaaggggag ggcaagccct acgagggcac
      901 ccagaccaac aggattaagg tggtggaggg aggccccctg ccgttcgcat tcgacatcct
      961 ggccacctgc tttatgtacg ggagcaagac cttcatcaac cacacccaag gaatcccaga
     1021 cttcttcaag caatccttcg taagtttaaa catgatttta ctaactaact aatctgattt
     1081 aaattttcag cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     1141 gcttaccgtt acccaggaca ccagcctcca ggacggctgc ttgatctaca acgtcaagct
     1201 cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     1261 ggccagtacc gagaccctgt accccgctga cggcggcctg gaaggcagat gcgacatggc
     1321 cctgaagctc gtgggcgggg gccacctgca ctgcaacctg aagaccacat acagatccaa
     1381 gaaacccgct aagaacctca agatgccagg agtaagttta aacatatata tactaactaa
     1441 ccctgattat ttaaattttc aggtctactt tgtggaccgc agactggaaa gaatcaagga
     1501 ggccgacaat gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct
     1561 ccctagcaaa ctggggcaca aacttaatgg catggacgag ctgtacaagg agggacgtgg
     1621 atccctcctc acctgcggag acgtcgagga gaacccagga ccaatgaccg ctccaaagaa
     1681 gaagcgcaag gttccagtcg agaagatgac tgccccaaaa aagaaacgta aggtcccgcg
     1741 ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc agcgcgctgt
     1801 gtcaaaagga gaagagctta ttaaggagaa tatgcgttcc aagctctacc tcgagggatc
     1861 cgtcaacgga caccaattca agtgcaccca cgagggagag ggaaagccat acgagggaaa
     1921 gcaaaccaac cgtatcaagg tcgtcgaggg aggaccactc ccattcgcct tcgacatcct
     1981 cgccacccac ttcatgtacg gatccaaggt aagtttaaac atatatatac taactaaccc
     2041 tgattattta aattttcagg tcttcatcaa gtacccagcc gacctcccag actacttcaa
     2101 gcaatccttc ccagagggat tcacctggga gcgtgtcatg gtcttcgagg acggaggagt
     2161 cctcaccgcc acccaagaca cctccctcca agacggagag ctcatctaca acgtcaaggt
     2221 ccgtggagtc aacttcccag ccaacggacc agtcatgcaa aagaagaccc tcggatggga
     2281 gccatccacc gagaccatgt acccagccga cggaggactc gagggacgtt gcgacaaggt
     2341 aagtttaaac atgattttac taactaacta atctgattta aattttcagg ccctcaagct
     2401 cgtcggagga ggacacctcc acgtcaactt caagaccacc tacaagtcca agaagccagt
     2461 caagatgcca ggagtccact acgtcgaccg tcgtctcgag cgtatcaagg aggccgacaa
     2521 cgagacctac gtcgagcaat acgagcacgc cgtcgcccgt tactccaacc tcggaggagg
     2581 aatggacgag ctctacaagc ctaggatacc ggtgggtgaa gacccaccaa agccatctgc
     2641 caagggagcc aagaaggccg ccaagaccgt tacgaagcca aaggacggaa agaagagacg
     2701 tcatgcccgt aaggaatcat actccgtcta catctaccgt gtcctcaagc aagttcatcc
     2761 agacactgga gtttcctcca aagccatgtc tatcatgaac tcttttgtca acgatgtctt
     2821 cgagcgtatt gctgctgaag catcccgtct tgctcactac aacaagcgtt ccacaatctc
     2881 atcccgcgaa attcagaccg ctgtccgtct gatccttcca ggagagcttg ccaagcacgc
     2941 cgtgtctgag ggaaccaagg ccgttaccaa gtacacttcc agcaagtagg aattccaact
     3001 gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc
     3061 atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat
     3121 ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt
     3181 tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt
     3241 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     3301 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     3361 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt
     3421 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     3481 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     3541 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     3601 attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga
     3661 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     3721 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     3781 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     3841 tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     3901 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3961 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4021 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4081 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     4141 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4261 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4321 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4381 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4441 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4501 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4561 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4621 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4681 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4741 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4801 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4861 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4921 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4981 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5041 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5101 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5161 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5221 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5281 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5341 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5401 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5461 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5521 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5581 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5641 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5701 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5761 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5821 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5881 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5941 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6001 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6061 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6121 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6181 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6241 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6301 agcggataac aatt
//
