LOCUS       sequence-382321-        8535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lim-4 fluorescent neural reporter driving nuclear mTagBFP2 
            expression (refer to NeuroPAL paper for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8535)
  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 8535)
  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: pEY22
FEATURES             Location/Qualifiers
     source          1..8535
                     /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             3900..3920
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3942..3962
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6126..6148)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6248..6267
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6345..6363)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6431..6535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6536..6604,6605..7396)
                     /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(6754..6773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7567..8155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8056..8075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8309..8326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8443..8464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8479..8484,8485..8502,8503..8509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8517..8533
                     /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(8522..8535,1..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 tgcctgcagc ttaaggtggc
      181 gaaggaacat ttgagaattc gagttgaatt agatgggcca tactgaactt taaaaataca
      241 gtgagatcat cggaaaccat gacgttagcc tcacgaacag tatacctttt ctgctgttct
      301 gccaatttgc atgcatttcg agaagtgaag gagaacttca acctgaaaat tagtgacaaa
      361 tattattttc ttattttcct cagactaatg caagattttg aaaaattgag tacgtaggcg
      421 taccccaaaa atgctcaacc acaataaata ttacggcgac ctacaagctt tgatttagaa
      481 attgtagttt cctaagccta aatgtacata ttacttttgg gttatgcctg agaattacca
      541 atgattgttt tgttcaataa gaattatcat ttacagagaa acaagatcca ttgcttttat
      601 cgattcgagt tgtgactttt taggataaaa tttccttgaa aaacgttaca gcgacagaca
      661 tcattttcga cgttgaatct gaaatcgtct gtaactcaag aaaaaatcgc ttccacgaaa
      721 ataaatttat ataaagttat tttcataaaa ttaactggga cttttgggtg attttgggag
      781 ttttcggcaa accatcaaat cgctaattga caaaataaca aaaaaggtat ttaaacgatt
      841 tttagataaa attcttaaat tttaaaacag tgaaactaaa acgggctggc atttatcctt
      901 gtagcgactc aacttaatat tagaacttga cttcacaatt taacagacac gacgtctgag
      961 gaactttttc ggttgtttgc atttttgata gctcactgtt ttatgcccat atacgtttta
     1021 tttcttattt atctcaaacc gtcttccctc aaacttgaat aaatcttcac aagacttttt
     1081 gggagtttct ggcggaccat caaatcgcta attgaattcg cgcattttcg tgttttgagt
     1141 gcgtccgcat ccttgcgagg ggaaatacga tttacatcga tctcgagata attcgattca
     1201 agctgttgca cacgtcctcc cgacaaacac tggagaagag cacccaccag ccaccgacgg
     1261 tcgacgactt ttgacttcaa attggatgtc aacaactttc tattaagcat gttttgaaat
     1321 gggatacaaa taaaagtaaa agaaagagat tcaaacatgg atattgaaaa aaagtaattc
     1381 agatatacac tataaagagg gcagactatt agtatggttt ttttcaaatg aaggagtgac
     1441 acggagtatc taaaaggtta acaaagttaa aggttaaaaa taacatttga aaaaagattt
     1501 aatctagaat tatacgaatt ttcaggtaat gcttttttga tgtgaaaatg caatttgaac
     1561 tctgaaaagt tggtaaatca aggttatttt attttagatc aaaactgagc ggcttgttcg
     1621 tcggctggta gatcaaatgt gaacttttta atgtttcagc acgtactgat ttggaaattg
     1681 ttacttcagt tgaataaaac tagtaaatca aaaaaaaatc taaaatttac catgtgttgg
     1741 cggaattgat ttattgtgat gtgtattgga ttttttgtgt attattgttt cttatgacac
     1801 gattcgtttg gaatcgtatt aaataaataa aaaataaaaa aaaaattgtc cacacgttca
     1861 agaaatacgt aattttaacc taccagccga cagccgacaa ccgacgtctg aacactataa
     1921 aaatatctaa atcgtggctg gttttttttg aagttttttt tctaagaagt atttcaagat
     1981 caaccaatca aaattttata attattctca aattgtcatt ttctgatact gtgaaaagta
     2041 aacaattttt taccagtggt ttagatttgc tgcatgtttt ttttttcatt tccagtcagt
     2101 ttctggagca aaataacgtc ctatatctct tcaggaagag gtttttcaga tttcaattcc
     2161 aatcttcaac cctcccacaa tcctaacaac taatagtatt aattttggta gaataataag
     2221 atgcttccaa tttgatgcac acgtgtcgag tggaatgcgc caaatggtgg tgtgtaggtg
     2281 tgcttcttcc ccgtctgttg ttctttttga gagcccccag agccaattca cattaatccg
     2341 ataactcttg ttatatatgt atgtgtatcg aatccgaaag gatcactatt ttaacaatag
     2401 tccttcggta tcactacctc attcatgttt tcctcagcgg catcctatga aaacggcagg
     2461 cctggcagat tgtgcgacat gaattgatgc tcattagcat gagacatcca agtcctcaag
     2521 actggtggca gtaacaagca aactagtcag gagcctccaa ttttggtggc tctattatct
     2581 ggccgaacaa tttaccctta ttgaaacaga tattagtcac cttttcctat tttttccgaa
     2641 aaacttggaa acgtaatatg ggatacgcag tagccatggt ttcattattt gagagagggg
     2701 ttgcaaaata tctctactta cttattatct tctttatgat aatattttgc ctccaaagcc
     2761 attatcacac atattttatt cgcaacggac acctaggatc atttttgaaa cttgtgtagt
     2821 ataaccccac attttgaata catagttcga atttattgta tagaaaaata caaatttcta
     2881 aggctgaaac gcaatatgtg tatcatttta attaaattgg atgaaattct agagttctaa
     2941 cataattttt aataaacagg attataatat tcaagtccca tattgccttg cacccaattg
     3001 tttaattttg tcagctttta tctcattttt agtttttttt tgcattttca ctgtattcta
     3061 gaggtggaca gcaattctac aaaattacct atgttccatt ttgttccaac tttttctatt
     3121 gaaaattgat attaaattaa gggaaattac ttaattagtt tgaaaatgac attcattgaa
     3181 acgttcaaga aaatgctgaa aattggttga ttttcaagta tcgaatagtc gaaatattta
     3241 atttttcagg acagtttaga gcacttttga aaacgtttca gctaaaataa atagattttt
     3301 tcggaaaaaa aaaccatctc aaaatatgca tttttgttca atggcctgga tgccagacca
     3361 tttcttaaaa tgtttaatgc tctaataaaa actatcaaac taattttttt tttgaaattt
     3421 agctgtttat ttattcatac ttcatatgat attaattatt ctcgatatta acactttatt
     3481 tgcacttgat aagcattaaa taatttccta ttgttataac tttaatttgg gaactttcag
     3541 attccggaga tctgggctcc ttcttttgcc ggacactcct acgcaagcag ccaaccacat
     3601 ccgccttgcc gcccaatagt gagcatccca gtgggcgggg cgagcttcta gtgaggcctt
     3661 ttcgacagaa caacagacta gaaggaagac ttttgcgtcg gtgctcgtcg actcgttagt
     3721 ggtccttgtt gtcttgttat ctacagctcg gccggcagtt cgttaagaaa gcagctgtca
     3781 caagttgcac ccgggattgg ccaaaggacc caaaggtatg tttcgaatga tactaacata
     3841 acatagaaca ttttcaggag gacccttgga gggtaccgag ctcagaaaaa atgaccgctc
     3901 caaagaagaa gcgcaaggtt ccagtcgaga agatgactgc cccaaaaaag aaacgtaagg
     3961 tcccgcggat gtcagagctt attaaggaga atatgcatat gaaactgtac atggaaggaa
     4021 ccgtggacaa ccatcacttt aagtgcactt cggaggggga aggcaaaccg tacgagggca
     4081 cccagaccat gcgaatcaag gtggtcgagg gaggcccact gcccttcgcc ttcgacattc
     4141 tggtaagttt aaacatatat atactaacta accctgatta tttaaatttt caggctacta
     4201 gcttcctgta cggcagcaag acttttatta atcacaccca gggcatcccg gacttcttca
     4261 aacagtcgtt ccctgagggc ttcacatggg agcgtgtgac cacatacgaa gacggtggcg
     4321 ttctcaccgc tacccaggac accagcctcc aggacggttg cctcatatat aatgtggtaa
     4381 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcagaag atccggggag
     4441 taaacttcac atccaatggt ccggttatgc agaagaaaac actcggctgg gaggccttca
     4501 ccgagacgct gtatccggct gacggaggcc tggaaggtcg aaacgacatg gccctgaagc
     4561 tcgtcggcgg tagccatctc atcgcaaacg ctaagaccac atatagatcc aagaaaccgg
     4621 taagtttaaa catgatttta ctaactaact aatctgattt aaattttcag gctaagaacc
     4681 ttaaaatgcc gggagtctac tatgttgact acagactgga acggatcaag gaggccaaca
     4741 acgagaccta cgtcgagcag cacgaggttg cagttgccag atactgcgac ctcccgagca
     4801 aactgggtca caagcttaat cctaggatac cggtgggtga agacccacca aagccatctg
     4861 ccaagggagc caagaaggcc gccaagaccg ttacgaagcc aaaggacgga aagaagagac
     4921 gtcatgcccg taaggaatca tactccgtct acatctaccg tgtcctcaag caagttcatc
     4981 cagacactgg agtttcctcc aaagccatgt ctatcatgaa ctcttttgtc aacgatgtct
     5041 tcgagcgtat tgctgctgaa gcatcccgtc ttgctcacta caacaagcgt tccacaatct
     5101 catcccgcga aattcagacc gctgtccgtc tgatccttcc aggagagctt gccaagcacg
     5161 ccgtgtctga gggaaccaag gccgttacca agtacacttc cagcaagtag gaattccaac
     5221 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     5281 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     5341 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     5401 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     5461 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     5521 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     5581 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     5641 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     5701 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     5761 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     5821 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     5881 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     5941 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     6001 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     6061 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6121 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6181 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6241 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     6301 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     6361 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6541 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6601 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6661 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6721 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6781 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6841 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6901 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6961 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7021 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7081 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7141 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7201 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7261 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7321 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7381 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7441 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7681 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8221 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     8281 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     8341 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     8401 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     8461 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     8521 gagcggataa caatt
//
