LOCUS       sequence-417731-        8395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mTurquoise2- and mVenus-inserted PcdhgB2 lacking an intracellular 
            domain (ICD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  AUTHORS   Hoshino N, Kanadome T, Takasugi T, Itoh M, Kaneko R, Inoue YU, Inoue
            T, Hirabayashi T, Watanabe M, Matsuda T, Nagai T, Tarusawa E, Yagi T
  TITLE     Visualization of trans homophilic interaction of clustered 
            protocadherin in neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Sep 19;120(38):e2301003120. doi: 
            10.1073/pnas.2301003120. Epub 2023 Sep 11.
  PUBMED    37695902
REFERENCE   2  (bases 1 to 8395)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-09-19"; volume: "120(38):e2301003120. 
            doi"; pages: " 10.1073/pnas.2301003120. Epub 2023 Sep 11"
COMMENT     Sequence Label: Combined gamma-B2-Delta-ICD-FRET probe
FEATURES             Location/Qualifiers
     source          1..8395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1934..1936,1937..2449)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(5417..5436)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5482..5537
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5536..5555)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5898..5914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5898..5914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5911..5933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5922..5938
                     /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(5946..5952,5953..5970,5971..5976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5991..6012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6071..6266
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6117..6252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6272..6406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6309..6328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6363..6382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6474..6491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6645..7233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6725..6744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7404..8195,8196..8264))
                     /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     8027..8046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8265..8369)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccagggggg gggggggggg gggagggggg
      541 ggggggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gaggaggggc tctgggagcg gtccggcctg gtggctgcag gtactgtcgc ctttcctgtt
     1801 gcttttgttc cacggagctc tcgcggatca gatccgctac tccattcccg aggagctggc
     1861 caagaactcg atagtaggaa acctcgcaag ggatctgggg ctcagtgtcc aggctagcgg
     1921 tggcggtggt tccgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1981 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2041 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2101 ctggcccacc ctcgtgacca ccctgagctg gggcgtgcag tgcttcgccc gctaccccga
     2161 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2221 taccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2281 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2341 cctggggcac aagctggagt acaactactt cagcgacaac gtctatatca ccgccgacaa
     2401 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     2461 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2521 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     2581 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggcg ctagcaatct
     2641 gcccacccgg aagctaaggg tcagcgcaga gaaggattat tttactgtaa atccagagag
     2701 tggggactta cttgtgggtg acagaataga cagagagcag gtttgcggga agcagcctca
     2761 gtgtgttttg gagttcgata ttgtcgctga gaagccctta aatattttcc acgtagctgt
     2821 aattcttcag gatgtgaacg acaacgctcc tctattcaaa cagagtgagg ttgatttaaa
     2881 cattggagag tccaccagac caggtaaagc atttccattg gatccagccc tcgatttaga
     2941 tgctggttct aattctctgc aaaaatacta tctcactgac aatgaatact ttgaagtcac
     3001 ggagaaagag accccagatg gacgtaaata tcctgagttg attctaaaac gttttctgga
     3061 cagagaagag cataattttc accaactggt cctcacggct gtggatgggg gagatcctcc
     3121 ccgaagtggc accacccaga tccgaatcca agtcacagac atcaatgata acccgccaat
     3181 gttcagccag gatgtgttca gtgtcaccct tcgggaggat gtaccaccag gcttctccgt
     3241 gcttcaggtg acagccaccg accaggacga gggtgtcaat gcagaaatca cctacgcttt
     3301 tcacaatgtg gatgaacaag tggaacggat ttttaactta gacaagagaa caggagaaat
     3361 cacgacaaag gataatttgg atttcgagac agctaagagt tacaccctga atgtagaagc
     3421 aaaagatccc ggagacctag catcccattg tagtatccaa gtgaaaattc tggatgagaa
     3481 tgattgtgta ccggaagtaa ttgtgacgtc agtatttacc cccctaccag aagattctcc
     3541 actaggaaca gtcatcgcct tgataaaaac cagagacaga gactctggag aaaatggaga
     3601 cgtctactgc catgtgttgg gaaatgaagg gtttgtattg aaatcttctt cgaagaatta
     3661 ttacaaacta gtgacagaca gaactctaga ccgggaggcg ataccggaat acaacgtcac
     3721 cattgtagct gccgataggg gtaagccccc gctctcctct aacgtaatca tcactctcca
     3781 tatctcagat gtcaacgaca atgcaccggt ttttcaccag gcctcctact tggtccatgt
     3841 ggctgagaac aacccacctg gaacctccat tgctcaagtc agtgcttctg acccggattt
     3901 gggatccaat ggcctcattt cctactccat catagccagc gacctagagc caagggcact
     3961 atcggctagc ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4021 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4081 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4141 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     4201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4261 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4381 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     4441 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     4501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4561 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4621 gctggagttc gtgaccgccg ccgggatccc accccctgct agctccttcg tgtccgtgaa
     4681 ccaggacagt ggagtggtgt tcgcgcagcg cgccttcgac cacgagcagc tgcgctcctt
     4741 ccagctgaca ctacaggcgc gggaccacgg atcgcccaca ctcagtgcca atgtgagcat
     4801 gcgtgtgctg gtgggcgacc gcaacgacaa cgcgccacgc gtgctgtatc ccacacttga
     4861 gcccgacggc tctgcgctct tcgacatggt gccgcgcgct gccgagcccg gatacctggt
     4921 caccaaggtg gtggctgtgg atgcagactc cggacacaat gcctggctat cttaccacgt
     4981 gctgcaggcc agcgatcccg ggctcttcag cctgggactg cgcacaggcg aggtgcgcac
     5041 agcgcgtgcc ctgggagaca gggactcagc gcggcagcgc cttctggtcg ctgtgcgtga
     5101 tggtggacag ccaccgctct ccgctacagc cacgctgcac ctcatcttcg ctgacagcct
     5161 gcaggaggtc ctgccagacc tcagcgatga ccctctacca tctgaccctc agagtgagct
     5221 tcagttttac ctggtggtgg ccttggccct gatctcagta ctcttcctcc tgtcggtgat
     5281 tctggctatt gccctgcgtc tgagacactc ctccagatca gctgcctggg gctgctttca
     5341 acctggcctc agctctaacc ttgcgactgg agttggcgtt gccatgccag gtgccgaaga
     5401 tgatgtggtg tgaagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     5461 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     5521 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     5581 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5641 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     5701 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     5761 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     5821 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     5881 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5941 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6001 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6061 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6181 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6241 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     6301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     6421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7321 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7381 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7441 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7501 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7561 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7621 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7681 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7741 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7801 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7861 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7921 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7981 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8041 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8101 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8161 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8221 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8281 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8341 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
