LOCUS       sequence-376016-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  klp-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 6936)
  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 6936)
  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: pEY48
FEATURES             Location/Qualifiers
     source          1..6936
                     /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             1221..1241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1266..1286
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1308..1328
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2232..2285
                     /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             2295..2315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2337..2357
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4527..4549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4649..4668
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4746..4764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4832..4936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4937..5005,5006..5797)
                     /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(5155..5174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5968..6556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6457..6476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6710..6727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6844..6865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6880..6885,6886..6903,6904..6910)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6918..6934
                     /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(6923..6936,1..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 tggaaagttt
      181 ggtaagttgc caaaatgact agatactttt aaatatctaa cacgttttgg agtttgctac
      241 gatattcggg ttttttggat aattatattc ataagtaaaa aatcggataa atgctgtcaa
      301 agttgaaata aactatccga ggctttatca aaaaatcaat tgttttcaga aattctgtgc
      361 atttattcaa ctttttgttc tctgattcca atcttcaatc ttcattaaat acaacttcat
      421 acaaattttt cttcttcaca tcaaacaccg tttggtccgt ttcctttcgt cgcttggaga
      481 cctacatggc aacattgact ccacgtgtat gtagaagaag ggtcccattc tccacgcttt
      541 actacactct atactgaatt ttccatttat tttcttgtcc ttttcatttt aaaaacattt
      601 ttccgttcgg atttgtttac aaattttgtt ctctctgata gaagtattca attattatac
      661 ttttaatctg taaattaaaa aaaaaaacta aatccttttc tcatatcacg ggaacaataa
      721 atcatgagaa tgcgtacttg gtgcatttta tttgacgcgc gatatctctt atcgaaaact
      781 acagtaagag cttaaatatc tattgttatt tgagttctta ctgtagtttt cgctaagaga
      841 tatcgcgcgt caaataaaat gcaccaagta cacattctca tgatttattg ttcccgtaat
      901 accaaccaat tattgcgacg caataccttc atatacattt tttaatttta aaaatttttt
      961 ttaaaatata gttgagatcc ttgttgtagt acggtagtag tacgaagata ttccaatgtt
     1021 tctttaaaaa aattttccaa ccaaatttag ttctaaattt attagttgaa cttttcagaa
     1081 tacccgggat tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga
     1141 acattttcag gaggaccctt ggagggtacc ggttttccca gtcacgacgc gtggtaccga
     1201 gctcagaaaa aatgaccgct ccaaagaaga aacgcaaagt accgagctca gaaaaaatga
     1261 cggcacctaa aaagaaacgc aaagtaccgg tcgccgagaa gacagcccca aaaaagaagc
     1321 gtaaagttcc acgcgtcatg ggagtaagtt taaacagttc ggtactaact aaccatacat
     1381 atttaaattt tcagcgcgct gtctccaagg gagaggagct catcaaggag aacatgcaca
     1441 ccaagctcta catggaggga accgtcaaca accaccactt caagtgcacc cacgaagggg
     1501 agggcaagcc ctacgagggc acccagacca acaggattaa ggtggtggag ggaggccccc
     1561 tgccgttcgc attcgacatc ctggccacct gctttatgta cgggagcaag accttcatca
     1621 accacaccca aggaatccca gacttcttca agcaatcctt cgtaagttta aacatgattt
     1681 tactaactaa ctaatctgat ttaaattttc agcctgaggg cttcacatgg gagagagtca
     1741 ccacatacga agacgggggc gtgcttaccg ttacccagga caccagcctc caggacggct
     1801 gcttgatcta caacgtcaag ctcagagggg tgaacttccc atccaacggc cctgtgatgc
     1861 agaagaaaac actcggctgg gaggccagta ccgagaccct gtaccccgct gacggcggcc
     1921 tggaaggcag atgcgacatg gccctgaagc tcgtgggcgg gggccacctg cactgcaacc
     1981 tgaagaccac atacagatcc aagaaacccg ctaagaacct caagatgcca ggagtaagtt
     2041 taaacatata tatactaact aaccctgatt atttaaattt tcaggtctac tttgtggacc
     2101 gcagactgga aagaatcaag gaggccgaca atgagaccta cgtcgagcag cacgaggtgg
     2161 ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat ggcatggacg
     2221 agctgtacaa ggagggacgt ggatccctcc tcacctgcgg agacgtcgag gagaacccag
     2281 gaccaatgac cgctccaaag aagaagcgca aggttccagt cgagaagatg actgccccaa
     2341 aaaagaaacg taaggtcccg cgggtaagtt taaacagttc ggtactaact aaccatacat
     2401 atttaaattt tcagcgcgct gtgtcaaaag gagaagagct tattaaggag aatatgcgtt
     2461 ccaagctcta cctcgaggga tccgtcaacg gacaccaatt caagtgcacc cacgagggag
     2521 agggaaagcc atacgaggga aagcaaacca accgtatcaa ggtcgtcgag ggaggaccac
     2581 tcccattcgc cttcgacatc ctcgccaccc acttcatgta cggatccaag gtaagtttaa
     2641 acatatatat actaactaac cctgattatt taaattttca ggtcttcatc aagtacccag
     2701 ccgacctccc agactacttc aagcaatcct tcccagaggg attcacctgg gagcgtgtca
     2761 tggtcttcga ggacggagga gtcctcaccg ccacccaaga cacctccctc caagacggag
     2821 agctcatcta caacgtcaag gtccgtggag tcaacttccc agccaacgga ccagtcatgc
     2881 aaaagaagac cctcggatgg gagccatcca ccgagaccat gtacccagcc gacggaggac
     2941 tcgagggacg ttgcgacaag gtaagtttaa acatgatttt actaactaac taatctgatt
     3001 taaattttca ggccctcaag ctcgtcggag gaggacacct ccacgtcaac ttcaagacca
     3061 cctacaagtc caagaagcca gtcaagatgc caggagtcca ctacgtcgac cgtcgtctcg
     3121 agcgtatcaa ggaggccgac aacgagacct acgtcgagca atacgagcac gccgtcgccc
     3181 gttactccaa cctcggagga ggaatggacg agctctacaa gcctaggata ccggtgggtg
     3241 aagacccacc aaagccatct gccaagggag ccaagaaggc cgccaagacc gttacgaagc
     3301 caaaggacgg aaagaagaga cgtcatgccc gtaaggaatc atactccgtc tacatctacc
     3361 gtgtcctcaa gcaagttcat ccagacactg gagtttcctc caaagccatg tctatcatga
     3421 actcttttgt caacgatgtc ttcgagcgta ttgctgctga agcatcccgt cttgctcact
     3481 acaacaagcg ttccacaatc tcatcccgcg aaattcagac cgctgtccgt ctgatccttc
     3541 caggagagct tgccaagcac gccgtgtctg agggaaccaa ggccgttacc aagtacactt
     3601 ccagcaagta ggaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc
     3661 aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac taactaacga
     3721 gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt ccaattactc
     3781 ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt gttattatca
     3841 aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa
     3901 ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc
     3961 tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta
     4021 tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca cacaaaatac
     4081 cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc acgtctgggc
     4141 ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt tggaattttt
     4201 caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg gtttccccta
     4261 ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt attgatgagc
     4321 acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa
     4381 gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg acagaataca
     4441 ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc ctttcgtctc
     4501 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4561 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4621 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4681 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcggcct
     4741 taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4801 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5461 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5641 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5761 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6601 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6661 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6721 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6781 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6841 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6901 tcgtatgttg tgtggaattg tgagcggata acaatt
//
