LOCUS       sequence-376011-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  flp-3 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 6179)
  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 6179)
  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: pEY44
FEATURES             Location/Qualifiers
     source          1..6179
                     /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             1854..1874
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1899..1919
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1941..1961
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3770..3792)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3892..3911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3989..4007)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4075..4179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4180..4248,4249..5040)
                     /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(4398..4417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5211..5799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5700..5719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6087..6108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6123..6128,6129..6146,6147..6153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6161..6177
                     /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(6166..6179,1..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 tgcctgcaga ctagttccgc
      181 atggaataat agcccatctg ttaaccaagt cttacgtttt cttcgcgaac aaaggaaacg
      241 actactggca ctgaatttca gattttagcc agattaggtc agaaaacgag gcacactatt
      301 ttcaagatca aaattttaaa attcagtctt gctagttctt caaagaattg tttcggaaat
      361 gtcaatgagt gctcataatt aaacatgtgg agtgactttg gaaggtgcta gaatttccaa
      421 gaagctttga tttattagtt ttcgttttaa tttcaaaaca ctgttcaatc ttgagaacaa
      481 gattagttca acctacttaa tcatttctcg cgttctgaca agtgacgttt ttgacaaaaa
      541 ataatgttta attattcaaa aacagaaaat cccccctcct taaagctcag ttcattaatc
      601 agcttcgctt cacaaattcg acaaggtggc atctcaacca acaggcacca atcggattct
      661 gtcatttctt tcttttttga gaacaaatgt ctatctacgt tttgaactaa caccagtccc
      721 ggtttaaatt tcgtacctga caattatcaa tcaagactta catttgacta gctgtgcacg
      781 caaagattta ataattggcg gtgggctgtg tgagagttga tgtcacaaag ttacagtatc
      841 ccatcaaaaa aatgggttcg agtcgtcatt ctcgttatcc atctaggctt gaattatgat
      901 gttgaacatc aaaatttata ggtttttcga gagatcccac ctcttccgtt aaccgccgcc
      961 ccgtttcaac acaattcttt cagtttcaaa ttattaacct gaactttgaa aactaccacc
     1021 tacgcttctt ctactaattt cgagtgtctt tttagactca attatcttat ttcttttctt
     1081 agagtattcc aaaaatgtca catgacgatg cataaattga ggtgccaccg ttttttaata
     1141 gaaactctgg ccgcaataaa agagcaatag cattgttact tggcaatttg tttgaaggag
     1201 tgttgggcgg cggtagacgt cgtacgtcaa atcgtgcatt ttcgaagtct tttctcgaca
     1261 caatagtaat cgtttgacct gtgcattgca acatcaaggg accacttact ttttagaaac
     1321 ccgccccttt aaaccaaatt gccaacgcct ccaattttta cctgacaata gaatgtaccg
     1381 agtacttatt aaacaaagtt gttcgccaaa gctcagattt tcaataattt ttgagcaccg
     1441 gatttcaatt ttactcgttt cacatgaaca tttttctaac tgcacacagt atttacgcat
     1501 tcctgaccaa atttctaaaa ccccacacac ctcttccggc atgacaactg aaccgttaaa
     1561 attgttgaaa ttaagctcat ataaaaacat gcaaatatct cggcggttca caacctcact
     1621 tctacttctt cttgactaaa ttttccgtaa ctaattgtta tattttaagg taccccttgt
     1681 gtgttagctc ccgcgtaaca ccaccataac caccacccgg gattggccaa aggacccaaa
     1741 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
     1801 accggttttc ccagtcacga cgcgtggtac cgagctcaga aaaaatgacc gctccaaaga
     1861 agaaacgcaa agtaccgagc tcagaaaaaa tgacggcacc taaaaagaaa cgcaaagtac
     1921 cggtcgccga gaagacagcc ccaaaaaaga agcgtaaagt tccacgcgtc atgggagtaa
     1981 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcagcgc gctgtctcca
     2041 agggagagga gctcatcaag gagaacatgc acaccaagct ctacatggag ggaaccgtca
     2101 acaaccacca cttcaagtgc acccacgaag gggagggcaa gccctacgag ggcacccaga
     2161 ccaacaggat taaggtggtg gagggaggcc ccctgccgtt cgcattcgac atcctggcca
     2221 cctgctttat gtacgggagc aagaccttca tcaaccacac ccaaggaatc ccagacttct
     2281 tcaagcaatc cttcgtaagt ttaaacatga ttttactaac taactaatct gatttaaatt
     2341 ttcagcctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctta
     2401 ccgttaccca ggacaccagc ctccaggacg gctgcttgat ctacaacgtc aagctcagag
     2461 gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcca
     2521 gtaccgagac cctgtacccc gctgacggcg gcctggaagg cagatgcgac atggccctga
     2581 agctcgtggg cgggggccac ctgcactgca acctgaagac cacatacaga tccaagaaac
     2641 ccgctaagaa cctcaagatg ccaggagtaa gtttaaacat atatatacta actaaccctg
     2701 attatttaaa ttttcaggtc tactttgtgg accgcagact ggaaagaatc aaggaggccg
     2761 acaatgagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     2821 gcaaactggg gcacaaactt aatggcatgg acgagctgta caagtaagcg ccggtcgcta
     2881 ccattaccaa cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta
     2941 aactgagttc tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct
     3001 ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc
     3061 accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt
     3121 ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa
     3181 aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca
     3241 caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat
     3301 agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt
     3361 tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt
     3421 tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct
     3481 tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc
     3541 taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga
     3601 ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt
     3661 tttgccttaa atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc
     3721 ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3781 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3841 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3901 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     3961 gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat
     4021 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4081 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4141 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4201 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4261 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4321 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4381 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4441 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4501 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4561 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4621 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4681 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4741 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4801 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4861 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4921 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4981 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5041 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5101 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5161 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5221 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5281 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5341 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5401 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5461 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5521 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5581 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5641 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5701 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5761 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5821 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5881 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5941 acgaccgagc gcagcgagtc agtgagcgat gaagcggaag agcgcccaat acgcaaaccg
     6001 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6061 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6121 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaatt
//
