LOCUS       sequence-376437-        7992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flp-20 fluorescent neural reporter driving nuclear CYOFP expression 
            (refer to NeuroPAL paper for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7992)
  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 7992)
  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: pEY28
FEATURES             Location/Qualifiers
     source          1..7992
                     /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             3351..3371
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3393..3413
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5583..5605)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5705..5724
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5802..5820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5888..5992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5993..6061,6062..6853)
                     /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(6211..6230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7024..7612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7513..7532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7766..7783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7900..7921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7936..7941,7942..7959,7960..7966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7974..7990
                     /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(7979..7992,1..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 aaacattggt
      181 cgggagatga tgaagatgac gaaaacgacg acctctcctt atgaatctat gagttcttgt
      241 catgaaacac gccaattttt atctcacata tgaatgtaga aactgagatc cacccccgac
      301 aagaaacccg aaaagtatta gtggatccat tttttagagg cgcagcaatt tcagcattta
      361 ctgttaagaa actgtaattc ttttaatcat ctaccgttat aaatcataat tcacctaatg
      421 gcctcacata cctaacttta tagttgccac tatgttcaac aacaacaaaa gtaaaaacaa
      481 tctgtaaact attccgaaaa gaagaaaaca atgtggtttt tgaattatgt gacaaatttt
      541 aataatcaaa gttaggaatt gggaaaaagc aaacaatttc gttacataat gtttagagca
      601 acgatactct tttttgattt gtaatacata gagcagctct agaaaacagt gatttgctga
      661 cttataaaga tctacttttt taatagttta cattgtgcag ctgttacggt ctgagatcta
      721 tctgagtcaa agttctggaa gaagttaaaa aactggtttt attattctat ttctttttgt
      781 tcttctcgtt tttggtttta aaaattattt ctctgatgtg tcaagtcttc tgaatcataa
      841 cgctacaact cttttagtca aaattatcaa aaattaaaaa aaaaggaatt aaaaaatttt
      901 tatatttcac agcaatcaat ttttagtttt gtcacaaggg taaaactgtg tttaacttct
      961 aaaaaacaca aataatttgt aaggaagtag tagcgccagt agagttgtct aaatacactt
     1021 acaatagtcg cagtgaacca aatacagaag ctcttttttt caagttttct ccgttatgaa
     1081 tatcatatta tcaaacccct tagacaagta atattgaaat tgtattcaaa tgaaagtttt
     1141 gcaacctagt gttttgatta ttaaaacgag agttaagtaa tctataactt caaggatacg
     1201 aatttataac attgttatgc gatttctgta gttttaacat cttcattcta gattctagac
     1261 atgttttcag tttccagtca taatttctat cgcgctctat caaattcaag ttaacatctt
     1321 cattggtcac cggaaagacg acggcccgcc taagttttta tgactaattc gaaaagtcgt
     1381 ttcggcttgt gccctcctac catcatactt cgctttttgg gactactggc aacaacgcca
     1441 gtgttgaggc gatatagaac ccatcatttt gaagatccga caacgacctt gttttttatt
     1501 tttccgactt tttcttcttc tatttcgaac ttttcagctt ttttgaaatg gcaatagcaa
     1561 cgaaaaaccc attttcatca caattatctc gaacttttag cagattactt ttccattacc
     1621 agtcactttc aaaattggaa aattcttaga aaatcagagg gtgactattc tgaaaacatt
     1681 attgtctttt gccgactatg cgctacatcc gattcatttt ttgtgagctc tttaagttat
     1741 tggtgttccc tcaatgttat cagttacaag acaaaccagt agtgttaaaa aacgccagtt
     1801 aacagttttc aacaaaaatc acaaaatgta acttgtttga gaaaacttgt aaaacggaaa
     1861 atactcaatt tgtatttaaa tatatgatat tatgtattaa aaatcatttt aagataaaca
     1921 gttcccatta tcttatgaaa attagtagat taaattcaaa ttaggctttc ttatatttct
     1981 ggaaagaaag agcttctgaa agtgttctga aaatcttatg atttgttcaa acatcaaagt
     2041 aaacatacaa acaaaataca catatttgtg ctaatgttct aaatatgcaa actttaaagt
     2101 taaagactaa agttgtttgc ccactctaat gtccataaga aataattcat acttatttta
     2161 ctagattatg agttctccaa ggcaattttt gatcgtgtat gattggtggc actctgaagt
     2221 tctaataata agataaagac aaaagataag gtttcttcct tttttaataa tcttatgaaa
     2281 aattgtttct agaacatctc tgaaagtatt ttattaaatc aaatgactgc tatagctgat
     2341 caatgaaaaa gtgctccccg tccaaaccgg aatataagta gttgtttcgc atggttcaac
     2401 ttcggtaccc aacctcataa aactaactac ttcttcactt tgtctcattt tggtgcattt
     2461 gaaaacgttg aactcggaca tgtattgccc atagtttaaa tttttgaata agctattttg
     2521 agatttaaaa cgttattttg gcattttatt tcaattttta tttaagtata ttcttgagtt
     2581 attcatgcat ttcatttttg atcactgaaa cccgattctg attcagatac aaaaactgaa
     2641 tttcgtgggt ttcgtgatat ttcacgattt gaagatttaa agattttgag atcaaacgga
     2701 gaagacccta atatagactg cgatgaaaca aaataacgtc acggaattct tttaaaatga
     2761 atggaatccg gagcccgctt ccttcatagt ttctgttttg caactctcgt atcctccatc
     2821 accagacaac aaaaatgctt catttgaaat gaattcgtga acagtttttt gtttttagtt
     2881 tagaactgcc gctcccttca gctccaccct cccatcaaat aggtgtgctc gatgtgcctc
     2941 gggacattgg atttattttt gtatccttgc tcatttttca tatgctttat tctttgtcta
     3001 ctccgtgaca tttttcaata atgctttcat tggaagttgc agtgaagttt gagttccata
     3061 ggtccgataa aatcataatt aatgagttct aggaagtgtt ttacaacatt tcaaattcat
     3121 aaatgttctc tccctcttcg tgattcattt tttgcataat tacgcttgat gattttttgt
     3181 tgttatgtgg ctcaatatgt tgtataaacc tttttttcag aaactttagt gagcaacccg
     3241 ggattggcca aaggacccaa aggtatgttt cgaatgatac taacataaca tagaacattt
     3301 tcaggaggac ccttgcttgg agggtaccga gctcagaaaa aatgaccgct ccaaagaaga
     3361 agcgcaaggt tccagtcgag aagatgactg ccccaaaaaa gaaacgtaag gtcccgcggg
     3421 taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag cgcgctgtgt
     3481 caaaaggaga agagcttatt aaggagaata tgcgttccaa gctctacctc gagggatccg
     3541 tcaacggaca ccaattcaag tgcacccacg agggagaggg aaagccatac gagggaaagc
     3601 aaaccaaccg tatcaaggtc gtcgagggag gaccactccc attcgccttc gacatcctcg
     3661 ccacccactt catgtacgga tccaaggtaa gtttaaacat atatatacta actaaccctg
     3721 attatttaaa ttttcaggtc ttcatcaagt acccagccga cctcccagac tacttcaagc
     3781 aatccttccc agagggattc acctgggagc gtgtcatggt cttcgaggac ggaggagtcc
     3841 tcaccgccac ccaagacacc tccctccaag acggagagct catctacaac gtcaaggtcc
     3901 gtggagtcaa cttcccagcc aacggaccag tcatgcaaaa gaagaccctc ggatgggagc
     3961 catccaccga gaccatgtac ccagccgacg gaggactcga gggacgttgc gacaaggtaa
     4021 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcaggcc ctcaagctcg
     4081 tcggaggagg acacctccac gtcaacttca agaccaccta caagtccaag aagccagtca
     4141 agatgccagg agtccactac gtcgaccgtc gtctcgagcg tatcaaggag gccgacaacg
     4201 agacctacgt cgagcaatac gagcacgccg tcgcccgtta ctccaacctc ggaggaggaa
     4261 tggacgagct ctacaagcct aggataccgg tgggtgaaga cccaccaaag ccatctgcca
     4321 agggagccaa gaaggccgcc aagaccgtta cgaagccaaa ggacggaaag aagagacgtc
     4381 atgcccgtaa ggaatcatac tccgtctaca tctaccgtgt cctcaagcaa gttcatccag
     4441 acactggagt ttcctccaaa gccatgtcta tcatgaactc ttttgtcaac gatgtcttcg
     4501 agcgtattgc tgctgaagca tcccgtcttg ctcactacaa caagcgttcc acaatctcat
     4561 cccgcgaaat tcagaccgct gtccgtctga tccttccagg agagcttgcc aagcacgccg
     4621 tgtctgaggg aaccaaggcc gttaccaagt acacttccag caagtaggaa ttccaactga
     4681 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
     4741 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
     4801 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
     4861 tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt
     4921 tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt
     4981 aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat
     5041 cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt
     5101 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     5161 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     5221 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     5281 taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg
     5341 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     5401 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     5461 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     5521 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5581 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5641 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5701 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5761 agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt
     5821 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5881 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5941 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6001 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6061 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6121 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6181 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6241 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6301 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6361 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6421 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6481 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6541 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6601 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6661 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6721 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6781 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6841 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6901 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6961 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7021 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7081 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7141 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7201 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7261 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7321 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7381 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7441 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7501 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7561 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7621 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7681 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7741 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7801 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7861 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7921 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7981 cggataacaa tt
//
