LOCUS       sequence-409847-        8995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP-A and ddGFP-B3-inserted Pcdh-alpha-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8995)
  AUTHORS   Kanadome T, Hoshino N, Nagai T, Yagi T, Matsuda T
  TITLE     Visualization of trans-interactions of a protocadherin-alpha between
            processes originating from single neurons.
  JOURNAL   iScience. 2023 Jul 17;26(7):107238. doi: 10.1016/j.isci.2023.107238.
            eCollection 2023 Jul 21.
  PUBMED    37534169
REFERENCE   2  (bases 1 to 8995)
  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: "iScience.";
            date: "2023-07-17"; volume: "26(7):107238. doi"; pages: " 
            10.1016/j.isci.2023.107238. eCollection 2023 Jul 21"
COMMENT     Sequence Label: pCX:cIPAD-3 (full length)
FEATURES             Location/Qualifiers
     source          1..8995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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"
     regulatory      1733..1742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5984..6010
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6017..6036)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6082..6137
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6136..6155)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6498..6514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6498..6514)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6511..6533)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6522..6538
                     /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(6546..6552,6553..6570,6571..6576))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6591..6612
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6671..6866
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6717..6852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6872..7006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6909..6928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6963..6982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7074..7091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7245..7833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7325..7344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8004..8795,8796..8864))
                     /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     8627..8646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8865..8969)
                     /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 gggggggggc gcsggggggg gggggggggg gggggggggg
      541 ggggggggcg 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 ggrggggcgc 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 ggaattttcc tggggaagtg gccaggaatc ccagcgcttg cttctttctt ttctgcttct
     1801 tgcaatctgg gaggcaggga acagcgctag cgcaaccatc aaagaattta tgaggttcaa
     1861 agtacgcatg gagggtagta tgaacgggca tgagttcgaa attgaaggag aaggggaagg
     1921 ccgcccctat gaggggaccc aaaccgccaa acttaaagtg acaaagggag gtccacttcc
     1981 ttttgcctgg gatatattga gcccacagtt catgtacgga agcgaagcct atgtacgaca
     2041 tcctgcagac ataccagatt acaagaagct gccctttcca gaaggcttta agtgggaaag
     2101 agtgatgaac tttgaagatg gcggcttggt aacagtgact caggacagca gtctccaaga
     2161 tggaactttg atctgtaagg tcaaaatgag gggtactaac tttccccccg atgggcccgt
     2221 catgcaaaaa aagacaatgg gctgggaagc ctcaaccgag atgttgtacc cagaagatgg
     2281 agtgctcaag ggacattctt atcaggcatt gaagctgaaa gatggcggac attacttggt
     2341 tgagtttgag actatctata tggccaagaa gcccgtacaa cttcctggag attactgcgt
     2401 agacaccaaa ctggatataa cctctcataa cgaggattac accattgtcg agcagtacgg
     2461 ccgcagtgaa ggtagacata gacttggaat ggatgagcga tataaggcta gccagatcca
     2521 ctactccatc cctgaggagg ccaaacacgg caccttcgtg ggccgcatcg cgcaggacct
     2581 ggggctggag ctgacggagc tggtgccccg cctgttcaga gtggcgtcca aggaccgcgg
     2641 agaccttctg gaggtaaatc tgcagaatgg cattttgttt gtgaactctc ggatcgaccg
     2701 ggaggagctg tgcgggcgga gcgcggagtg cagcatccac ctggaggtga tcgtggacag
     2761 gccgttgcag gttttccacg tggaggtgga ggtgagggac attaacgaca accctcccag
     2821 gttcccaaca acacaaaaga atctgttcat tgcagaatca aggccacttg acacttggtt
     2881 tccactagag ggcgcttcag acgcagatat cggaatcaat gctgtactga cttacagact
     2941 gagtccaaat gattactttt ctttggaaaa accatccaac gacgaacggg taaaaggtct
     3001 tggacttgta ttacggaaat ctttagaccg ggaggaaact ccagagataa ttttagtgct
     3061 tactgtcacg gacggaggaa agccagagct gaccggcagt gttcagttac tcatcactgt
     3121 gctggatgcc aatgataatg ctccagtttt tgacagatcc ctgtataccg tgaaattacc
     3181 agaaaacgtt ccaaatggga cattggtagt caaagtcaat gcctcagatt tagacgaagg
     3241 ggtaaatggg gatattatgt actcattttc tacagatatt tcaccaaatg tgaaatacaa
     3301 attccacata gaccctgtta gcggagagat tattgtaaag ggatacattg attttgaaga
     3361 atgcaaatcc tatgaaattc tcatagaggg aattgacaag ggacaacttc cactctctgg
     3421 gcactgtaaa gtcattgtac aagttgaaga catcaacgat aatgttccag aattggaatt
     3481 taaatctcta tcacttccaa tacgagagaa ctctccagtg ggcactgtca tcgcactcat
     3541 tagtgtgtct gatcgggaca cgggtgtcaa cgggcaggtg acctgctccc tgacaagtca
     3601 tgctagcgtt agtaagagcg aggaggtaat caaggagttt atgaggttta aggtcaggct
     3661 cgagggcagc atgaatgggc atgagtttga gatagagggt gaaggggaag gcagaccata
     3721 tgagggtact caaacagcca agctcaaagt aaccaagggc ggtccactcc ctttcgcatg
     3781 ggatattttg agtcccctta ttatgtatgg ttccaaaatg tatgtgaagc accctgcaga
     3841 tgtgcctgat tatatgaaac tgtcattccc tgagggtttc aagtgggaaa gagtgatgca
     3901 ttttgaagat ggtggactcg tcacagctac acaggacagt tccttgcaag atggtacact
     3961 tatttacaaa gtaaagatgc ggggtacaaa cttccctcca gatggtcccg ttatgcagaa
     4021 gaagacactc ggttgggact acgcaacaga gcgactctat cccgaggagg gcgttctcaa
     4081 aggtgaactg ctgggacgac tgaaactcaa ggacggcgga ctgaatctgg tggaaagtaa
     4141 gaccatatac atggctaaaa aacccgtaca actcccagga tactattttg tcgacacaaa
     4201 gctggatatt acctcccata atgaggatta caccattgta gagcaatacg agaggagtga
     4261 aggccggcat catctgggac atggtacagg ctctactgga tctggcagta gcgctagcgt
     4321 ccccttcaag ttggtgtcca cattcaagaa ttactattcg ctcgtgctgg acagcgccct
     4381 ggacagagag acaacagcgg actataaggt ggtggtgaca gcgcgggatg ggggctctcc
     4441 ctcgctgtgg gccacggcta gtgtgtctgt tgaggttgct gacgtgaacg acaatgcacc
     4501 tgtgttcgcg cagcccgaat acacggtgtt cgtgaaggag aacaacccgc ctggtgcgca
     4561 catcttcacg gtgtcagcga tggatgcgga cgcacaggag aacgcgctgg tgtcctactc
     4621 gctggtggag cggagggtgg gcgagcgctt gctgtcgagc tatgtgtctg tgcacgcgga
     4681 gagcggcaag gtgttcgcgc tgcagcctct ggaccatgag gagctggagc tgctgcggtt
     4741 ccaggtgagc gcgcgggatg ctggtgtacc tgccctgggc agcaatgtga ctctgcaggt
     4801 gtttgtgctg gacgagaatg acaacgcgcc cacactgctg gaacctgagg caggagtctc
     4861 tggtggaatc gtgagccggt tggtgtccag atcagtgggt gcaggccatg tggtggctaa
     4921 ggtgcgcgcg gtggatgcag actctggcta taatgcatgg ctctcttatg agctgcaatc
     4981 gtcagaaggc aattcccgta gccttttccg cgtaggtttg tatacgggcg agattagtac
     5041 tacgcgcata ctggatgaag cagattcgcc acgtcagcgc cttctggtgc tggtgaagga
     5101 ccatggtgac ccagcaatga ttgttaccgc cacagtgttg gtgtctctgg tagagaatgg
     5161 cccggtacca aaggctccat cgcgagtatc cacgagtgtc acacactctg aggcgtcact
     5221 ggtggatgtc aacgtgtacc tgatcattgc catctgtgca gtgtccagcc tgctagtgct
     5281 cacgctgctg ctgtacacag cgctgcgctg ttccactgtc cccagtgaga gcgtgtgcgg
     5341 gcctccaaaa ccggtaatgg tgtgctccag tgcagtgggg agctggtcat actcccaaca
     5401 aaggaggcaa agggtgtgct ctggggagta cccacctaag accgacctca tggccttcag
     5461 ccccagttta tctgattcaa gggacagaga ggatcaattg cagtctgcag aggattcctc
     5521 tggaaagccc cggcagccca accctgactg gcgctactct gcctcgctaa gagcaggcat
     5581 gcacagctct gtgcacctgg aggaggctgg cattctacgg gctggtccag gagggcctga
     5641 tcagcagtgg ccaacagtat ccagtgcaac accagaacct gaggcaggag aggtgtcccc
     5701 tccggtgggc gccggtgtca acagcaacag ctggaccttt aaatacggac caggcaaccc
     5761 caaacagtcc ggtcccggtg agttgccaga caaattcatt atcccaggat ctcctgcaat
     5821 catctccatc cggcaggagc ctgctaacaa ccaaattgac aaaagcgatt ttataacctt
     5881 cggcaaaaag gaggagacca agaaaaagaa gaaaaagaag aagggtaaca agacccagga
     5941 gaaaaaagag aaagggaaca gcacgacgga caacagtgac cagtatccgt acgacgtacc
     6001 ggattatgcg tgaagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     6061 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     6121 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     6181 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     6241 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     6301 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     6361 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     6421 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     6481 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6661 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6841 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     6901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6961 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     7021 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7081 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7141 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7201 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7261 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7321 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7381 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7441 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7501 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7561 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7621 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7681 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7741 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7801 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7861 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7921 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7981 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8041 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8101 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8161 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8221 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8281 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8341 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8401 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8461 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8521 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8581 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8641 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8701 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8761 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8821 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8881 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8941 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
