LOCUS       sequence-409849-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ddGFP-A-inserted Pcdh-alpha-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8308)
  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 8308)
  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:alpha-4-EC4-GA
FEATURES             Location/Qualifiers
     source          1..8308
                     /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             5297..5323
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5330..5349)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5395..5450
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5449..5468)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5811..5827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5811..5827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5824..5846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5835..5851
                     /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(5859..5865,5866..5883,5884..5889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5904..5925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5984..6179
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6030..6165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6185..6319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6222..6241)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6276..6295
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6387..6404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6558..7146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6638..6657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7317..8108,8109..8177))
                     /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     7940..7959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8178..8282)
                     /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 gggggggsgs gccrgggggg 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 ggggccgcck 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 acagccagat ccactactcc atccctgagg aggccaaaca
     1861 cggcaccttc gtgggccgca tcgcgcagga cctggggctg gagctgacgg agctggtgcc
     1921 ccgcctgttc agagtggcgt ccaaggaccg cggagacctt ctggaggtaa atctgcagaa
     1981 tggcattttg tttgtgaact ctcggatcga ccgggaggag ctgtgcgggc ggagcgcgga
     2041 gtgcagcatc cacctggagg tgatcgtgga caggccgttg caggttttcc acgtggaggt
     2101 ggaggtgagg gacattaacg acaaccctcc caggttccca acaacacaaa agaatctgtt
     2161 cattgcagaa tcaaggccac ttgacacttg gtttccacta gagggcgctt cagacgcaga
     2221 tatcggaatc aatgctgtac tgacttacag actgagtcca aatgattact tttctttgga
     2281 aaaaccatcc aacgacgaac gggtaaaagg tcttggactt gtattacgga aatctttaga
     2341 ccgggaggaa actccagaga taattttagt gcttactgtc acggacggag gaaagccaga
     2401 gctgaccggc agtgttcagt tactcatcac tgtgctggat gccaatgata atgctccagt
     2461 ttttgacaga tccctgtata ccgtgaaatt accagaaaac gttccaaatg ggacattggt
     2521 agtcaaagtc aatgcctcag atttagacga aggggtaaat ggggatatta tgtactcatt
     2581 ttctacagat atttcaccaa atgtgaaata caaattccac atagaccctg ttagcggaga
     2641 gattattgta aagggataca ttgattttga agaatgcaaa tcctatgaaa ttctcataga
     2701 gggaattgac aagggacaac ttccactctc tgggcactgt aaagtcattg tacaagttga
     2761 agacatcaac gataatgttc cagaattgga atttaaatct ctatcacttc caatacgaga
     2821 gaactctcca gtgggcactg tcatcgcact cattagtgtg tctgatcggg acacgggtgt
     2881 caacgggcag gtgacctgct ccctgacaag tcatgctagc gttagtaaga gcgaggaggt
     2941 aatcaaggag tttatgaggt ttaaggtcag gctcgagggc agcatgaatg ggcatgagtt
     3001 tgagatagag ggtgaagggg aaggcagacc atatgagggt actcaaacag ccaagctcaa
     3061 agtaaccaag ggcggtccac tccctttcgc atgggatatt ttgagtcccc ttattatgta
     3121 tggttccaaa atgtatgtga agcaccctgc agatgtgcct gattatatga aactgtcatt
     3181 ccctgagggt ttcaagtggg aaagagtgat gcattttgaa gatggtggac tcgtcacagc
     3241 tacacaggac agttccttgc aagatggtac acttatttac aaagtaaaga tgcggggtac
     3301 aaacttccct ccagatggtc ccgttatgca gaagaagaca ctcggttggg actacgcaac
     3361 agagcgactc tatcccgagg agggcgttct caaaggtgaa ctgctgggac gactgaaact
     3421 caaggacggc ggactgaatc tggtggaaag taagaccata tacatggcta aaaaacccgt
     3481 acaactccca ggatactatt ttgtcgacac aaagctggat attacctccc ataatgagga
     3541 ttacaccatt gtagagcaat acgagaggag tgaaggccgg catcatctgg gacatggtac
     3601 aggctctact ggatctggca gtagcgctag cgtccccttc aagttggtgt ccacattcaa
     3661 gaattactat tcgctcgtgc tggacagcgc cctggacaga gagacaacag cggactataa
     3721 ggtggtggtg acagcgcggg atgggggctc tccctcgctg tgggccacgg ctagtgtgtc
     3781 tgttgaggtt gctgacgtga acgacaatgc acctgtgttc gcgcagcccg aatacacggt
     3841 gttcgtgaag gagaacaacc cgcctggtgc gcacatcttc acggtgtcag cgatggatgc
     3901 ggacgcacag gagaacgcgc tggtgtccta ctcgctggtg gagcggaggg tgggcgagcg
     3961 cttgctgtcg agctatgtgt ctgtgcacgc ggagagcggc aaggtgttcg cgctgcagcc
     4021 tctggaccat gaggagctgg agctgctgcg gttccaggtg agcgcgcggg atgctggtgt
     4081 acctgccctg ggcagcaatg tgactctgca ggtgtttgtg ctggacgaga atgacaacgc
     4141 gcccacactg ctggaacctg aggcaggagt ctctggtgga atcgtgagcc ggttggtgtc
     4201 cagatcagtg ggtgcaggcc atgtggtggc taaggtgcgc gcggtggatg cagactctgg
     4261 ctataatgca tggctctctt atgagctgca atcgtcagaa ggcaattccc gtagcctttt
     4321 ccgcgtaggt ttgtatacgg gcgagattag tactacgcgc atactggatg aagcagattc
     4381 gccacgtcag cgccttctgg tgctggtgaa ggaccatggt gacccagcaa tgattgttac
     4441 cgccacagtg ttggtgtctc tggtagagaa tggcccggta ccaaaggctc catcgcgagt
     4501 atccacgagt gtcacacact ctgaggcgtc actggtggat gtcaacgtgt acctgatcat
     4561 tgccatctgt gcagtgtcca gcctgctagt gctcacgctg ctgctgtaca cagcgctgcg
     4621 ctgttccact gtccccagtg agagcgtgtg cgggcctcca aaaccggtaa tggtgtgctc
     4681 cagtgcagtg gggagctggt catactccca acaaaggagg caaagggtgt gctctgggga
     4741 gtacccacct aagaccgacc tcatggcctt cagccccagt ttatctgatt caagggacag
     4801 agaggatcaa ttgcagtctg cagaggattc ctctggaaag ccccggcagc ccaaccctga
     4861 ctggcgctac tctgcctcgc taagagcagg catgcacagc tctgtgcacc tggaggaggc
     4921 tggcattcta cgggctggtc caggagggcc tgatcagcag tggccaacag tatccagtgc
     4981 aacaccagaa cctgaggcag gagaggtgtc ccctccggtg ggcgccggtg tcaacagcaa
     5041 cagctggacc tttaaatacg gaccaggcaa ccccaaacag tccggtcccg gtgagttgcc
     5101 agacaaattc attatcccag gatctcctgc aatcatctcc atccggcagg agcctgctaa
     5161 caaccaaatt gacaaaagcg attttataac cttcggcaaa aaggaggaga ccaagaaaaa
     5221 gaagaaaaag aagaagggta acaagaccca ggagaaaaaa gagaaaggga acagcacgac
     5281 ggacaacagt gaccagtatc cgtacgacgt accggattat gcgtgaagat ctttttccct
     5341 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     5401 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     5461 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     5521 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     5581 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     5641 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     5701 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5761 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     5821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5941 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     6001 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6061 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6121 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     6181 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6301 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6361 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6421 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6481 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6541 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6601 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6661 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6721 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6781 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6841 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6901 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6961 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7021 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7081 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7141 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7201 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7261 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7321 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7381 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7441 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7501 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7561 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7621 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7681 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7741 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7801 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7861 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7921 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7981 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8041 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8101 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8161 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8221 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8281 cgcacatttc cccgaaaagt gccacctg
//
