LOCUS       sequence-375933-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gcy-21 fluorescent neural reporter driving nuclear CyOFP1 and 
            mTagBFP2 expression (refer to NeuroPAL paper for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  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 7759)
  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: pEY47
FEATURES             Location/Qualifiers
     source          1..7759
                     /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             1747..1767
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1789..1809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3061..3114
                     /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             3124..3144
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3166..3186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5350..5372)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5472..5491
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5569..5587)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5655..5759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5760..5828,5829..6620)
                     /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(5978..5997)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6791..7379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7280..7299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7533..7550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7667..7688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7703..7708,7709..7726,7727..7733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7741..7757
                     /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(7746..7759,1..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 aaactgggag
      181 tgaaagcatc tccggcgatg ggctcgacga gcattatgtg atgttgttat gcctttgaat
      241 aaatttgttc ccattttaaa tgcaaaatta atagttaaaa attgttggaa atgtttaaaa
      301 aaattttcag ttatattcgg tatggtgtac gtggtgtaag agagaaaaaa attcccacat
      361 ttttttgtag atcaaaccgt aatgggacat cctgacacca catgcatatc gggactggat
      421 tctatcattt gggtcaaaaa aacccacaga cgatactcca ccgttaggta ccttgcgcga
      481 aaccggtgaa acatcggctg ctaatcacac atcatatggg aaatgacaaa ttaaaatgca
      541 acgatacaaa atactgtgtt tagcagccga catgtcgctg gttccagctg ggagggagag
      601 cgcagagaag catgcacttt agctgtctgc gccctatcta aaactacaaa ttactctgtt
      661 agctttgcag ttaggcatac caaaagtaca tttttttaga atcatcaaga aaaactgatc
      721 cggatatgat atttctgcaa atatttctag cctaaacttt ttcttttcac ctgttttgga
      781 tcaccttttt tcaggaggcg gagtagattt tccagatttt tgcctttttt tgttacttgc
      841 aaacttatgt aaatattttt gtgtttctct aaaattgcct atcgactttt ccatttcttc
      901 ctggctaaga atctactgct atattgtggc catagcttca tttcctttaa actttgggac
      961 tattatgctt ataatttata agaatgataa aatcgataat tttaagtatt tcatgctttg
     1021 gtttcaggtt actttgaatc tttaattttc aatttaatca aggtcaaatt tcagataacc
     1081 aactcaatta ccgacttata tatggcaatt ttcgtccaac caatgtctct tatttgaata
     1141 tttggtcaaa ttatttgttt gtaagaatta aagtttataa cctaccattt aagaaattcc
     1201 cattacagtc tcttctccca gcactagtcg ccgtgcaatt tgtcagcatt taattccgcg
     1261 aaagatgcca cctacctctt taataatagt ttctctcact tgacatgttt cttattaatt
     1321 ttccaataaa attaactatt ccagcaaaaa atgagttttg ggtactgcat gggtactcta
     1381 tgggtgcagg gttactgtat ttatgcaata taagggtaca ccgtacaaaa ataaaattca
     1441 aattttcatt ttcgagttct caatatacat ttcatttccg gtgatggtgg gtgttgacgc
     1501 atcaattttc taatttccgg ttccaagctc aacctaaagt catgcctaat tccgcactaa
     1561 tcctttgtaa tttccggtta tccgggaaaa aaccattgtt ctaaataaat ttcattttca
     1621 tattattctg ctcccgggat tggccaaagg acccaaaggt atgtttcgaa tgatactaac
     1681 ataacataga acattttcag gaggaccctt gcttggaggg taccgagctc agaaaaaatg
     1741 accgctccaa agaagaagcg caaggttcca gtcgagaaga tgactgcccc aaaaaagaaa
     1801 cgtaaggtcc cgcgggtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat
     1861 tttcagcgcg ctgtgtcaaa aggagaagag cttattaagg agaatatgcg ttccaagctc
     1921 tacctcgagg gatccgtcaa cggacaccaa ttcaagtgca cccacgaggg agagggaaag
     1981 ccatacgagg gaaagcaaac caaccgtatc aaggtcgtcg agggaggacc actcccattc
     2041 gccttcgaca tcctcgccac ccacttcatg tacggatcca aggtaagttt aaacatatat
     2101 atactaacta accctgatta tttaaatttt caggtcttca tcaagtaccc agccgacctc
     2161 ccagactact tcaagcaatc cttcccagag ggattcacct gggagcgtgt catggtcttc
     2221 gaggacggag gagtcctcac cgccacccaa gacacctccc tccaagacgg agagctcatc
     2281 tacaacgtca aggtccgtgg agtcaacttc ccagccaacg gaccagtcat gcaaaagaag
     2341 accctcggat gggagccatc caccgagacc atgtacccag ccgacggagg actcgaggga
     2401 cgttgcgaca aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     2461 caggccctca agctcgtcgg aggaggacac ctccacgtca acttcaagac cacctacaag
     2521 tccaagaagc cagtcaagat gccaggagtc cactacgtcg accgtcgtct cgagcgtatc
     2581 aaggaggccg acaacgagac ctacgtcgag caatacgagc acgccgtcgc ccgttactcc
     2641 aacctcggag gaggaatgga cgagctctac aagcctagga taccggtggg tgaagaccca
     2701 ccaaagccat ctgccaaggg agccaagaag gccgccaaga ccgttacgaa gccaaaggac
     2761 ggaaagaaga gacgtcatgc ccgtaaggaa tcatactccg tctacatcta ccgtgtcctc
     2821 aagcaagttc atccagacac tggagtttcc tccaaagcca tgtctatcat gaactctttt
     2881 gtcaacgatg tcttcgagcg tattgctgct gaagcatccc gtcttgctca ctacaacaag
     2941 cgttccacaa tctcatcccg cgaaattcag accgctgtcc gtctgatcct tccaggagag
     3001 cttgccaagc acgccgtgtc tgagggaacc aaggccgtta ccaagtacac ttccagcaag
     3061 gagggacgtg gatccctcct cacctgcgga gacgtcgagg agaacccagg accaatgacc
     3121 gctccaaaga agaagcgcaa ggttccagtc gagaagatga ctgccccaaa aaagaaacgt
     3181 aaggtcccgc ggatgtcaga gcttattaag gagaatatgc atatgaaact gtacatggaa
     3241 ggaaccgtgg acaaccatca ctttaagtgc acttcggagg gggaaggcaa accgtacgag
     3301 ggcacccaga ccatgcgaat caaggtggtc gagggaggcc cactgccctt cgccttcgac
     3361 attctggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcaggct
     3421 actagcttcc tgtacggcag caagactttt attaatcaca cccagggcat cccggacttc
     3481 ttcaaacagt cgttccctga gggcttcaca tgggagcgtg tgaccacata cgaagacggt
     3541 ggcgttctca ccgctaccca ggacaccagc ctccaggacg gttgcctcat atataatgtg
     3601 gtaagtttaa acagttcggt actaactaac catacatatt taaattttca gaagatccgg
     3661 ggagtaaact tcacatccaa tggtccggtt atgcagaaga aaacactcgg ctgggaggcc
     3721 ttcaccgaga cgctgtatcc ggctgacgga ggcctggaag gtcgaaacga catggccctg
     3781 aagctcgtcg gcggtagcca tctcatcgca aacgctaaga ccacatatag atccaagaaa
     3841 ccggtaagtt taaacatgat tttactaact aactaatctg atttaaattt tcaggctaag
     3901 aaccttaaaa tgccgggagt ctactatgtt gactacagac tggaacggat caaggaggcc
     3961 aacaacgaga cctacgtcga gcagcacgag gttgcagttg ccagatactg cgacctcccg
     4021 agcaaactgg gtcacaagct taatcctagg ataccggtgg gtgaagaccc accaaagcca
     4081 tctgccaagg gagccaagaa ggccgccaag accgttacga agccaaagga cggaaagaag
     4141 agacgtcatg cccgtaagga atcatactcc gtctacatct accgtgtcct caagcaagtt
     4201 catccagaca ctggagtttc ctccaaagcc atgtctatca tgaactcttt tgtcaacgat
     4261 gtcttcgagc gtattgctgc tgaagcatcc cgtcttgctc actacaacaa gcgttccaca
     4321 atctcatccc gcgaaattca gaccgctgtc cgtctgatcc ttccaggaga gcttgccaag
     4381 cacgccgtgt ctgagggaac caaggccgtt accaagtaca cttccagcaa gtaggaattc
     4441 caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata ataggggccg
     4501 ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat ttaaattttc
     4561 agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg
     4621 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt
     4681 tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa
     4741 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata
     4801 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat
     4861 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt
     4921 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat
     4981 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt
     5041 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc
     5101 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     5161 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     5221 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     5281 ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga
     5341 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5401 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5461 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5521 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     5581 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5941 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6001 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6061 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6301 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7441 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7501 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7561 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7621 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7681 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7741 ttgtgagcgg ataacaatt
//
