LOCUS       sequence-464051-        7538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of inhibitory post-synapse using GPHN-FingR-mGold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7538)
  AUTHORS   Lee J, Lai S, Yang S, Zhao S, Blanco FA, Lyons AC, Merino-Urteaga R,
            Ahrens JF, Nguyen NA, Liu H, Liu Z, Lambert GG, Shaner NC, Chen L, 
            Tolias KF, Zhang J, Ha T, St-Pierre F
  TITLE     Bright and photostable yellow fluorescent proteins for extended 
            imaging.
  JOURNAL   Nat Commun. 2025 Apr 4;16(1):3241. doi: 10.1038/s41467-025-58223-5.
  PUBMED    40185748
REFERENCE   2  (bases 1 to 7538)
  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"; journalName: "Nat 
            Commun."; date: "2025-04-4"; volume: "16(1):3241. doi"; pages: " 
            10.1038/s41467-025-58223-5"
COMMENT     Sequence Label: pCAG-GPHN-FingR-mGold-CCR5TC
FEATURES             Location/Qualifiers
     source          1..7538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    25..36
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        40..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          698..1714
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1638..1660
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1722..1741
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2062..2085
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2110..2112,2113..2820)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(2152..2173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2413..2432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2760..2781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2866..3207
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     CDS             3256..3381
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     intron          3430..3562
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3545..3564
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3571..4159
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3624..3644)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4042..4053)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4186..4203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4192..4303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4399..4419)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4399..4417)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4438..4454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4560..4579)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4625..4680
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4679..4698)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5041..5057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5065..5081
                     /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)."
     promoter        complement(join(5089..5095,5096..5113,5114..5119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5134..5155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5214..5409
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5260..5395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5415..5549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5506..5525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5617..5634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5788..6376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5868..5887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6547..7338,7339..7407))
                     /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     7170..7189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7408..7512)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagagtcatc ctcatcgtcg acattgatta ttgactagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      481 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      541 nsgcgcgccr gggggggggg gggggggsgg gggggggggs ggggsgrggs ggrgrggkgc
      601 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      661 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      721 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      781 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
      841 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg
      901 ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      961 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
     1021 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
     1081 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     1141 tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca
     1201 cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg
     1261 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcckcggg ccggggrggg
     1321 ctcgggggrg gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
     1381 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1441 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     1501 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     1561 ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg
     1621 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     1681 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     1741 gctgtctcat cattttggca aagaattcgg taccgcgggc ccgggattac aatgctcgaa
     1801 gtcaaggaag catcaccaac cagcatccag atcagctggg gcaagtacaa ggtcatggtt
     1861 cgctactacc gcatcaccta cggtgaaact ggtggcaata gccctgtcca ggaattcacc
     1921 gtgcctggca gcaagtccac tgctaccatc agcagcctga aacctggtgt cgactatacc
     1981 atcacggtgt acgccgtcac gatcgaccac tggaactacc aggacccgat cccgatctcc
     2041 atcaactacc gcaccggatc cgattacaag gatgacgacg ataagggtag cggctccagt
     2101 agatctgggg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2161 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2221 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2281 cccaccctcg tgaccagcct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     2341 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2401 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2461 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2521 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     2581 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     2641 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2701 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2761 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2821 tctaggggag ctggcgctgg agcgggtgca ggggctggct ctagattcca gtgccggatc
     2881 tgcatgcgga acttcagcga ccggtccaac ctgagcaggc acatcagaac ccacaccgga
     2941 gaaaagccct tcgcctgcga catttgcggc cggaagttcg ccatcagcag caacctgaac
     3001 agccacacca agatccacac tggcagccag aaacctttcc agtgcagaat ttgtatgaga
     3061 aactttagca gaagcgacaa cctggccaga cacatccgga cacatactgg tgaaaaacct
     3121 tttgcctgtg atatctgtgg cagaaagttt gccacctccg gcaatctgac ccggcacaca
     3181 aagattcacc tgcggggcag ccagctatcg attgtcgacg ctcctgaaca acgtgaaggt
     3241 gcttctcaag tttctgtttc tgttactttt gaagatgttg ctgttctttt tactcgtgat
     3301 gaatggaaaa aacttgatct ttctcaacgt tctctttatc gtgaagttat gcttgaaaat
     3361 tattctaatc ttgcttctat ggcttaaacg cgttgtacaa gtaaagcggc cccggactcg
     3421 aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     3481 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     3541 cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa aatttgtgaa
     3601 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3661 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3721 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3781 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3841 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3901 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3961 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4021 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4081 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4141 ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg actgtgcctt
     4201 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4261 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4321 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4381 atagcccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     4441 atagctgttt cctgtgtgaa attgttatcc gctaattcac tcctcaggtg caggctgcct
     4501 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcgat
     4561 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     4621 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     4681 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     4741 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     4801 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     4861 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     4921 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     4981 gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     5041 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5101 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5161 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     5221 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5281 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5341 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5401 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     5581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7501 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
