LOCUS       sequence-220264-        6739 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6739)
  AUTHORS   Gross GG, Junge JA, Mora RJ, Kwon HB, Olson CA, Takahashi TT, Liman 
            ER, Ellis-Davies GC, McGee AW, Sabatini BL, Roberts RW, Arnold DB
  TITLE     Recombinant probes for visualizing endogenous synaptic proteins in 
            living neurons.
  JOURNAL   Neuron. 2013 Jun 19;78(6):971-85. doi: 10.1016/j.neuron.2013.04.017.
  PUBMED    23791193
REFERENCE   2  (bases 1 to 6739)
  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.neuron.2013.04"; journalName: "Neuron"; date: 
            "2013-06-19- 19"; volume: "78"; issue: "6"; pages: "971-85"
COMMENT     Sequence Label: pCAG_GPHN.FingR-eGFP-CCR5TC
FEATURES             Location/Qualifiers
     source          1..6739
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     321..343
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     405..424
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             745..768
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(790..792,793..1500)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     CDS             join(790..792,793..1500)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(832..853)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1093..1112)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1440..1461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1546..1887
                     /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             1936..2061
                     /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          2110..2242
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2225..2244
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2251..2839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2304..2324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2722..2733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2866..2883)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2872..2983
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3079..3099)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3079..3097)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3214..3233)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3279..3334
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3333..3352)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3695..3711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3695..3711)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3708..3730)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3719..3735
                     /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(3743..3749,3750..3767,3768..3773))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3788..3809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3868..4063
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3914..4049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4069..4203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4106..4125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4160..4179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4271..4288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4442..5030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4522..4541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5201..5992,5993..6061))
                     /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     5824..5843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6062..6166)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    6216..6227
                     /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        6231..6610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
       61 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      121 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      181 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
      241 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
      301 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
      361 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
      421 tgtgctgtct catcattttg gcaaagaatt cggtaccgcg ggcccgggat tacaatgctc
      481 gaagtcaagg aagcatcacc aaccagcatc cagatcagct ggggcaagta caaggtcatg
      541 gttcgctact accgcatcac ctacggtgaa actggtggca atagccctgt ccaggaattc
      601 accgtgcctg gcagcaagtc cactgctacc atcagcagcc tgaaacctgg tgtcgactat
      661 accatcacgg tgtacgccgt cacgatcgac cactggaact accaggaccc gatcccgatc
      721 tccatcaact accgcaccgg atccgattac aaggatgacg acgataaggg tagcggctcc
      781 agatctgggg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      841 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      901 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      961 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1021 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1081 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1141 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1201 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1261 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1321 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1381 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1441 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1501 tctaggggag ctggcgctgg agcgggtgca ggggctggct ctagattcca gtgccggatc
     1561 tgcatgcgga acttcagcga ccggtccaac ctgagcaggc acatcagaac ccacaccgga
     1621 gaaaagccct tcgcctgcga catttgcggc cggaagttcg ccatcagcag caacctgaac
     1681 agccacacca agatccacac tggcagccag aaacctttcc agtgcagaat ttgtatgaga
     1741 aactttagca gaagcgacaa cctggccaga cacatccgga cacatactgg tgaaaaacct
     1801 tttgcctgtg atatctgtgg cagaaagttt gccacctccg gcaatctgac ccggcacaca
     1861 aagattcacc tgcggggcag ccagctatcg attgtcgacg ctcctgaaca acgtgaaggt
     1921 gcttctcaag tttctgtttc tgttactttt gaagatgttg ctgttctttt tactcgtgat
     1981 gaatggaaaa aacttgatct ttctcaacgt tctctttatc gtgaagttat gcttgaaaat
     2041 tattctaatc ttgcttctat ggcttaaacg cgttgtacaa gtaaagcggc cccggactcg
     2101 aggccgcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     2161 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     2221 cactttgcct ttctctccac aggtgtcgac aatcaacctc tggattacaa aatttgtgaa
     2281 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2341 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2401 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2461 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2521 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2581 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2641 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2701 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2761 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2821 ctttgggccg cctccccgcc tggaattcga gctcggtacg atcagcctcg actgtgcctt
     2881 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2941 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3001 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3061 atagcccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3121 atagcykaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     3181 aatgccctgg ctcacaaata ccactgagat cgatcttttt ccctctgcca aaaattatgg
     3241 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3301 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3361 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3421 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3481 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3541 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3601 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     3661 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     3721 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3781 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3841 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     3901 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3961 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4021 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     4081 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4141 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4201 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6181 aagtgccacc tggtcgacat tgattattga ctagagtcat cctcatcgtc gacattgatt
     6241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     6361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     6421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     6481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6601 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     6661 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     6721 gggggggggg gcgcgcgcc
//
