LOCUS       sequence-464047-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of inhibitory post-synapse using GPHN-FingR-mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  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 7537)
  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-mVenus-CCR5TC
FEATURES             Location/Qualifiers
     source          1..7537
                     /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          697..1713
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1637..1659
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1721..1740
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2061..2084
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2109..2111,2112..2819)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(2151..2172)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2412..2431)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2759..2780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2865..3206
                     /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             3255..3380
                     /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          3429..3561
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3544..3563
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3570..4158
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3623..3643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4041..4052)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4185..4202)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4191..4302
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4398..4418)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4398..4416)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4437..4453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4559..4578)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4624..4679
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4678..4697)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5040..5056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5064..5080
                     /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(5088..5094,5095..5112,5113..5118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5133..5154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5213..5408
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5259..5394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5414..5548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5451..5470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5505..5524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5616..5633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5787..6375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5867..5886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6546..7337,7338..7406))
                     /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     7169..7188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7407..7511)
                     /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 sgcgcgccrg gggggggggg gggggggggg gggsggggsg gggsgrggsg grgrggkgcg
      601 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      661 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      721 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      781 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      841 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      901 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      961 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1021 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1081 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1141 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1201 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1261 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgccycgggc cggggrgggc
     1321 tcgggggrgg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1381 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1441 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1501 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1561 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1621 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1681 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1741 ctgtctcatc attttggcaa agaattcggt accgcgggcc cgggattaca atgctcgaag
     1801 tcaaggaagc atcaccaacc agcatccaga tcagctgggg caagtacaag gtcatggttc
     1861 gctactaccg catcacctac ggtgaaactg gtggcaatag ccctgtccag gaattcaccg
     1921 tgcctggcag caagtccact gctaccatca gcagcctgaa acctggtgtc gactatacca
     1981 tcacggtgta cgccgtcacg atcgaccact ggaactacca ggacccgatc ccgatctcca
     2041 tcaactaccg caccggatcc gattacaagg atgacgacga taagggtagc ggctccagta
     2101 gatctggggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2161 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2221 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     2281 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     2341 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2401 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2461 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2521 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     2581 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     2641 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2701 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     2761 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtact
     2821 ctaggggagc tggcgctgga gcgggtgcag gggctggctc tagattccag tgccggatct
     2881 gcatgcggaa cttcagcgac cggtccaacc tgagcaggca catcagaacc cacaccggag
     2941 aaaagccctt cgcctgcgac atttgcggcc ggaagttcgc catcagcagc aacctgaaca
     3001 gccacaccaa gatccacact ggcagccaga aacctttcca gtgcagaatt tgtatgagaa
     3061 actttagcag aagcgacaac ctggccagac acatccggac acatactggt gaaaaacctt
     3121 ttgcctgtga tatctgtggc agaaagtttg ccacctccgg caatctgacc cggcacacaa
     3181 agattcacct gcggggcagc cagctatcga ttgtcgacgc tcctgaacaa cgtgaaggtg
     3241 cttctcaagt ttctgtttct gttacttttg aagatgttgc tgttcttttt actcgtgatg
     3301 aatggaaaaa acttgatctt tctcaacgtt ctctttatcg tgaagttatg cttgaaaatt
     3361 attctaatct tgcttctatg gcttaaacgc gttgtacaag taaagcggcc ccggactcga
     3421 ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     3481 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     3541 actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa
     3601 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3661 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3721 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3781 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3841 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3901 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3961 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4021 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4081 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4141 tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga ctgtgccttc
     4201 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4261 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4321 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4381 tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4441 tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc aggctgccta
     4501 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgatc
     4561 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4621 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4681 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4741 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4801 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4861 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4921 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4981 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     5041 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5101 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5161 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5221 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5281 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5341 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5401 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5461 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5521 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7501 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
