LOCUS       sequence-409845-        7366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EBFP2-fused Pcdh-gamma-B2 lacking an EC1 domain and an ICD domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  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 7366)
  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:gamma-B2-Delta-EC1-Delta-ICD-EB
FEATURES             Location/Qualifiers
     source          1..7366
                     /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"
     CDS             join(3668..3670,3671..4384)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIKV
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLLE
                     FRTAAGITLGMDELYK"
     primer_bind     complement(3710..3731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4388..4407)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4453..4508
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4507..4526)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4869..4885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4869..4885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4882..4904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4893..4909
                     /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(4917..4923,4924..4941,4942..4947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4962..4983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5042..5237
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5088..5223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5243..5377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5280..5299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5334..5353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5445..5462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5616..6204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5696..5715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6375..7166,7167..7235))
                     /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     6998..7017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7236..7340)
                     /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 rggsggagag 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 tcgcggacaa cgctcctcta ttcaaacaga gtgaggttga
     1861 tttaaacatt ggagagtcca ccagaccagg taaagcattt ccattggatc cagccctcga
     1921 tttagatgct ggttctaatt ctctgcaaaa atactatctc actgacaatg aatactttga
     1981 agtcacggag aaagagaccc cagatggacg taaatatcct gagttgattc taaaacgttt
     2041 tctggacaga gaagagcata attttcacca actggtcctc acggctgtgg atgggggaga
     2101 tcctccccga agtggcacca cccagatccg aatccaagtc acagacatca atgataaccc
     2161 gccaatgttc agccaggatg tgttcagtgt cacccttcgg gaggatgtac caccaggctt
     2221 ctccgtgctt caggtgacag ccaccgacca ggacgagggt gtcaatgcag aaatcaccta
     2281 cgcttttcac aatgtggatg aacaagtgga acggattttt aacttagaca agagaacagg
     2341 agaaatcacg acaaaggata atttggattt cgagacagct aagagttaca ccctgaatgt
     2401 agaagcaaaa gatcccggag acctagcatc ccattgtagt atccaagtga aaattctgga
     2461 tgagaatgat tgtgtaccgg aagtaattgt gacgtcagta tttacccccc taccagaaga
     2521 ttctccacta ggaacagtca tcgccttgat aaaaaccaga gacagagact ctggagaaaa
     2581 tggagacgtc tactgccatg tgttgggaaa tgaagggttt gtattgaaat cttcttcgaa
     2641 gaattattac aaactagtga cagacagaac tctagaccgg gaggcgatac cggaatacaa
     2701 cgtcaccatt gtagctgccg ataggggtaa gcccccgctc tcctctaacg taatcatcac
     2761 tctccatatc tcagatgtca acgacaatgc accggttttt caccaggcct cctacttggt
     2821 ccatgtggct gagaacaacc cacctggaac ctccattgct caagtcagtg cttctgaccc
     2881 ggatttggga tccaatggcc tcatttccta ctccatcata gccagcgacc tagagccaag
     2941 ggcactatcg tccttcgtgt ccgtgaacca ggacagtgga gtggtgttcg cgcagcgcgc
     3001 cttcgaccac gagcagctgc gctccttcca gctgacacta caggcgcggg accacggatc
     3061 gcccacactc agtgccaatg tgagcatgcg tgtgctggtg ggcgaccgca acgacaacgc
     3121 gccacgcgtg ctgtatccca cacttgagcc cgacggctct gcgctcttcg acatggtgcc
     3181 gcgcgctgcc gagcccggat acctggtcac caaggtggtg gctgtggatg cagactccgg
     3241 acacaatgcc tggctatctt accacgtgct gcaggccagc gatcccgggc tcttcagcct
     3301 gggactgcgc acaggcgagg tgcgcacagc gcgtgccctg ggagacaggg actcagcgcg
     3361 gcagcgcctt ctggtcgctg tgcgtgatgg tggacagcca ccgctctccg ctacagccac
     3421 gctgcacctc atcttcgctg acagcctgca ggaggtcctg ccagacctca gcgatgaccc
     3481 tctaccatct gaccctcaga gtgagcttca gttttacctg gtggtggcct tggccctgat
     3541 ctcagtactc ttcctcctgt cggtgattct ggctattgcc ctgcgtctga gacactcctc
     3601 cagatcagct gcctggggct gctttcaacc tggcctcagc tctaaccttg cgactggagt
     3661 tgctagcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3721 ggacggcgac gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac
     3781 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3841 caccctcgtg accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat
     3901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3961 cttcttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac
     4021 cctggtgaac cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg
     4081 gcacaagctg gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa
     4141 gaacggcatc aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct
     4201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag
     4261 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     4321 ggtcctgctg gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4381 gtaaagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     4441 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     4501 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     4561 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     4621 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     4681 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     4741 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4801 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     4861 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4921 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4981 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     5041 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5161 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5221 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     5281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5341 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5401 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6361 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6421 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6481 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6541 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6601 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6661 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6721 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6781 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6841 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6901 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6961 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7021 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7081 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7141 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7201 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7261 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7321 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
