LOCUS       sequence-409830-        8278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP-B3-inserted Pcdh-alpha-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8278)
  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 8278)
  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:B3-alpha-4
FEATURES             Location/Qualifiers
     source          1..8278
                     /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             5267..5293
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5300..5319)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5365..5420
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5419..5438)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5781..5797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5781..5797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5794..5816)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5805..5821
                     /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(5829..5835,5836..5853,5854..5859))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5874..5895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5954..6149
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6000..6135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6155..6289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6192..6211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6246..6265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6357..6374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6528..7116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6608..6627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7287..8078,8079..8147))
                     /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     7910..7929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8148..8252)
                     /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 gggggggsgc gccrgggggg gggggggggg gggggggggg
      541 ggggggggsg rggcggagag 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 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 tgtccccttc aagttggtgt ccacattcaa gaattactat tcgctcgtgc tggacagcgc
     3661 cctggacaga gagacaacag cggactataa ggtggtggtg acagcgcggg atgggggctc
     3721 tccctcgctg tgggccacgg ctagtgtgtc tgttgaggtt gctgacgtga acgacaatgc
     3781 acctgtgttc gcgcagcccg aatacacggt gttcgtgaag gagaacaacc cgcctggtgc
     3841 gcacatcttc acggtgtcag cgatggatgc ggacgcacag gagaacgcgc tggtgtccta
     3901 ctcgctggtg gagcggaggg tgggcgagcg cttgctgtcg agctatgtgt ctgtgcacgc
     3961 ggagagcggc aaggtgttcg cgctgcagcc tctggaccat gaggagctgg agctgctgcg
     4021 gttccaggtg agcgcgcggg atgctggtgt acctgccctg ggcagcaatg tgactctgca
     4081 ggtgtttgtg ctggacgaga atgacaacgc gcccacactg ctggaacctg aggcaggagt
     4141 ctctggtgga atcgtgagcc ggttggtgtc cagatcagtg ggtgcaggcc atgtggtggc
     4201 taaggtgcgc gcggtggatg cagactctgg ctataatgca tggctctctt atgagctgca
     4261 atcgtcagaa ggcaattccc gtagcctttt ccgcgtaggt ttgtatacgg gcgagattag
     4321 tactacgcgc atactggatg aagcagattc gccacgtcag cgccttctgg tgctggtgaa
     4381 ggaccatggt gacccagcaa tgattgttac cgccacagtg ttggtgtctc tggtagagaa
     4441 tggcccggta ccaaaggctc catcgcgagt atccacgagt gtcacacact ctgaggcgtc
     4501 actggtggat gtcaacgtgt acctgatcat tgccatctgt gcagtgtcca gcctgctagt
     4561 gctcacgctg ctgctgtaca cagcgctgcg ctgttccact gtccccagtg agagcgtgtg
     4621 cgggcctcca aaaccggtaa tggtgtgctc cagtgcagtg gggagctggt catactccca
     4681 acaaaggagg caaagggtgt gctctgggga gtacccacct aagaccgacc tcatggcctt
     4741 cagccccagt ttatctgatt caagggacag agaggatcaa ttgcagtctg cagaggattc
     4801 ctctggaaag ccccggcagc ccaaccctga ctggcgctac tctgcctcgc taagagcagg
     4861 catgcacagc tctgtgcacc tggaggaggc tggcattcta cgggctggtc caggagggcc
     4921 tgatcagcag tggccaacag tatccagtgc aacaccagaa cctgaggcag gagaggtgtc
     4981 ccctccggtg ggcgccggtg tcaacagcaa cagctggacc tttaaatacg gaccaggcaa
     5041 ccccaaacag tccggtcccg gtgagttgcc agacaaattc attatcccag gatctcctgc
     5101 aatcatctcc atccggcagg agcctgctaa caaccaaatt gacaaaagcg attttataac
     5161 cttcggcaaa aaggaggaga ccaagaaaaa gaagaaaaag aagaagggta acaagaccca
     5221 ggagaaaaaa gagaaaggga acagcacgac ggacaacagt gaccagtatc cgtacgacgt
     5281 accggattat gcgtgaagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     5341 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     5401 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     5461 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     5521 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     5581 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     5641 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     5701 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     5761 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5821 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5881 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5941 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6061 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6121 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     6181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6241 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     6301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
