LOCUS       sequence-417743-        7696 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mTurquoise2-inserted PcdhgB2 lacking an intracellular domain (ICD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7696)
  AUTHORS   Kanadome T, Hoshino N, Nagai T, Matsuda T, Yagi T
  TITLE     Development of FRET-based indicators for visualizing homophilic 
            trans interaction of a clustered protocadherin.
  JOURNAL   Sci Rep. 2021 Nov 15;11(1):22237. doi: 10.1038/s41598-021-01481-2.
  PUBMED    34782670
REFERENCE   2  (bases 1 to 7696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2021-11-15"; volume: "11(1):22237. doi"; pages: " 
            10.1038/s41598-021-01481-2"
COMMENT     Sequence Label: pCX:gamma-B2-Delta-ICD-EC1-G4S-mTQ2-Delta-C6
FEATURES             Location/Qualifiers
     source          1..7696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1933..1935,1936..2448)
                     /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(1975..1996)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2236..2255)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2583..2604
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4717..4736)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4782..4837
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4836..4855)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5198..5214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5198..5214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5211..5233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5222..5238
                     /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(5246..5252,5253..5270,5271..5276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5291..5312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5371..5566
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5417..5552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5572..5706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5609..5628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5663..5682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5774..5791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6025..6044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6704..7495,7496..7564))
                     /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     7327..7346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7565..7669)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggsgsgsg csaggggggg gggggggggg sgaggggggg
      541 ggsggggsga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcckc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgccaccatg
     1741 aggaggggct ctgggagcgg tccggcctgg tggctgcagg tactgtcgcc tttcctgttg
     1801 cttttgttcc acggagctct cgcggatcag atccgctact ccattcccga ggagctggcc
     1861 aagaactcga tagtaggaaa cctcgcaagg gatctggggc tcagtgtcca ggctagcggt
     1921 ggcggtggtt ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1981 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2041 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2101 tggcccaccc tcgtgaccac cctgagctgg ggcgtgcagt gcttcgcccg ctaccccgac
     2161 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgt
     2221 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2281 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2341 ctggggcaca agctggagta caactacttc agcgacaacg tctatatcac cgccgacaag
     2401 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     2461 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2521 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     2581 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcgc tagcaatctg
     2641 cccacccgga agctaagggt cagcgcagag aaggattatt ttactgtaaa tccagagagt
     2701 ggggacttac ttgtgggtga cagaatagac agagagcagg tttgcgggaa gcagcctcag
     2761 tgtgttttgg agttcgatat tgtcgctgag aagcccttaa atattttcca cgtagctgta
     2821 attcttcagg atgtgaacga caacgctcct ctattcaaac agagtgaggt tgatttaaac
     2881 attggagagt ccaccagacc aggtaaagca tttccattgg atccagccct cgatttagat
     2941 gctggttcta attctctgca aaaatactat ctcactgaca atgaatactt tgaagtcacg
     3001 gagaaagaga ccccagatgg acgtaaatat cctgagttga ttctaaaacg ttttctggac
     3061 agagaagagc ataattttca ccaactggtc ctcacggctg tggatggggg agatcctccc
     3121 cgaagtggca ccacccagat ccgaatccaa gtcacagaca tcaatgataa cccgccaatg
     3181 ttcagccagg atgtgttcag tgtcaccctt cgggaggatg taccaccagg cttctccgtg
     3241 cttcaggtga cagccaccga ccaggacgag ggtgtcaatg cagaaatcac ctacgctttt
     3301 cacaatgtgg atgaacaagt ggaacggatt tttaacttag acaagagaac aggagaaatc
     3361 acgacaaagg ataatttgga tttcgagaca gctaagagtt acaccctgaa tgtagaagca
     3421 aaagatcccg gagacctagc atcccattgt agtatccaag tgaaaattct ggatgagaat
     3481 gattgtgtac cggaagtaat tgtgacgtca gtatttaccc ccctaccaga agattctcca
     3541 ctaggaacag tcatcgcctt gataaaaacc agagacagag actctggaga aaatggagac
     3601 gtctactgcc atgtgttggg aaatgaaggg tttgtattga aatcttcttc gaagaattat
     3661 tacaaactag tgacagacag aactctagac cgggaggcga taccggaata caacgtcacc
     3721 attgtagctg ccgatagggg taagcccccg ctctcctcta acgtaatcat cactctccat
     3781 atctcagatg tcaacgacaa tgcaccggtt tttcaccagg cctcctactt ggtccatgtg
     3841 gctgagaaca acccacctgg aacctccatt gctcaagtca gtgcttctga cccggatttg
     3901 ggatccaatg gcctcatttc ctactccatc atagccagcg acctagagcc aagggcacta
     3961 tcgtccttcg tgtccgtgaa ccaggacagt ggagtggtgt tcgcgcagcg cgccttcgac
     4021 cacgagcagc tgcgctcctt ccagctgaca ctacaggcgc gggaccacgg atcgcccaca
     4081 ctcagtgcca atgtgagcat gcgtgtgctg gtgggcgacc gcaacgacaa cgcgccacgc
     4141 gtgctgtatc ccacacttga gcccgacggc tctgcgctct tcgacatggt gccgcgcgct
     4201 gccgagcccg gatacctggt caccaaggtg gtggctgtgg atgcagactc cggacacaat
     4261 gcctggctat cttaccacgt gctgcaggcc agcgatcccg ggctcttcag cctgggactg
     4321 cgcacaggcg aggtgcgcac agcgcgtgcc ctgggagaca gggactcagc gcggcagcgc
     4381 cttctggtcg ctgtgcgtga tggtggacag ccaccgctct ccgctacagc cacgctgcac
     4441 ctcatcttcg ctgacagcct gcaggaggtc ctgccagacc tcagcgatga ccctctacca
     4501 tctgaccctc agagtgagct tcagttttac ctggtggtgg ccttggccct gatctcagta
     4561 ctcttcctcc tgtcggtgat tctggctatt gccctgcgtc tgagacactc ctccagatca
     4621 gctgcctggg gctgctttca acctggcctc agctctaacc ttgcgactgg agttggcgtt
     4681 gccatgccag gtgccgaaga tgatgtggtg tgaagatctt tttccctctg ccaaaaatta
     4741 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     4801 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4861 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     4921 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4981 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5041 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5101 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     5161 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5221 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5281 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5341 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5401 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5461 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5521 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     5581 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5641 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5701 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgg
//
