LOCUS       sequence-204179-        6384 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 6384)
  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 6384)
  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-mKate2-IL2RGTC
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     39..61
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     123..142
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             463..486
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             505..1197
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     CDS             1597..1722
                     /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          1765..1897
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1880..1899
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1906..2494
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1959..1979)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2377..2388)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2521..2538)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2527..2638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2734..2754)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2734..2752)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2773..2789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2891..2910)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2956..3011
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3010..3029)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3372..3388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3396..3412
                     /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(3420..3426,3427..3444,3445..3450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3465..3486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3545..3740
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3591..3726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3746..3880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3783..3802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3837..3856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3948..3965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4119..4707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4199..4218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4878..5669,5670..5738))
                     /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     5501..5520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5739..5843)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5968..6271
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
       61 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      121 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg gtaccgcggg
      181 cccgggatta caatgctcga agtcaaggaa gcatcaccaa ccagcatcca gatcagctgg
      241 ggcaagtaca aggtcatggt tcgctactac cgcatcacct acggtgaaac tggtggcaat
      301 agccctgtcc aggaattcac cgtgcctggc agcaagtcca ctgctaccat cagcagcctg
      361 aaacctggtg tcgactatac catcacggtg tacgccgtca cgatcgacca ctggaactac
      421 caggacccga tcccgatctc catcaactac cgcaccggat ccgattacaa ggatgacgac
      481 gataagggta gcggctccag atctgtgagc gagctgatta aggagaacat gcacatgaag
      541 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
      601 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
      661 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
      721 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
      781 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      841 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
      901 atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc
      961 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1021 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1081 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1141 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagatct
     1201 agatttcagt gccagatttg catgcgcaac tttagccgca aaagcaccct gaccgatcat
     1261 attcgcaccc ataccggcga aaaaccgttt gcgtgcgata tttgcggccg caaatttgcg
     1321 gcgcgcagca cccgcaccac ccataccaaa attcataccg gcagccagaa accgtttcag
     1381 tgccgcattt gcatgcgcaa ctttagccgc agcgatagcc tgagcaaaca tattcgcacc
     1441 cataccggcg aaaaaccgtt tgcgtgcgat atttgcggcc gcaaatttgc gcagcgcagc
     1501 aacctgaaag tgcataccaa aattcatctg cgcggcagcc agctgatcga tggtgtcgac
     1561 gctcctgaac aacgtgaagg tgcttctcaa gtttctgttt ctgttacttt tgaagatgtt
     1621 gctgttcttt ttactcgtga tgaatggaaa aaacttgatc tttctcaacg ttctctttat
     1681 cgtgaagtta tgcttgaaaa ttattctaat cttgcttcta tggcttagcg tacaagtaaa
     1741 gcggccccgg actcgaggcc gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     1801 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     1861 ggtcttactg acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat
     1921 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1981 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2041 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2101 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2161 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2221 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2281 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2341 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2401 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2461 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag
     2521 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2701 aggattggga agacaatagc ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     2761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc
     2821 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2881 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2941 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     3001 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3061 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3121 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3181 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3241 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3301 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     3361 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3421 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3481 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     3541 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3661 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3721 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     3781 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3841 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     3901 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3961 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4141 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4201 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4261 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4321 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4381 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4441 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4501 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4561 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4621 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4681 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4741 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4801 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4861 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4921 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4981 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5041 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5101 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5161 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5221 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5281 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5341 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5401 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5461 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5521 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5581 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5641 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5701 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5761 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5821 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtcgacattg
     5881 attattgact agtcttccac agagtactag ttattaatag taatcaatta cggggtcatt
     5941 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6001 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6061 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6121 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6181 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6241 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     6301 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     6361 ttgtgcagcg atgggggcgg gggg
//
