LOCUS       sequence-426464-        7491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses red #1.4 pre-SynapShot and TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7491)
  AUTHORS   Son S, Nagahama K, Lee J, Jung K, Kwak C, Kim J, Noh YW, Kim E, Lee 
            S, Kwon HB, Heo WD
  TITLE     Real-time visualization of structural dynamics of synapses in live 
            cells in vivo.
  JOURNAL   Nat Methods. 2024 Jan 8. doi: 10.1038/s41592-023-02122-4.
  PUBMED    38191933
REFERENCE   2  (bases 1 to 7491)
  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: "Nat 
            Methods. 2024 Jan 8. doi: 10.1038/s41592-023-02122-4."
COMMENT     Sequence Label: #1.4 pre-R-SynapShot-TagBFP
FEATURES             Location/Qualifiers
     source          1..7491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(159..747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(239..258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(918..1709,1710..1778))
                     /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     1541..1560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1779..1883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1909..2364)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2046..2067)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2258..2277
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2436..2565
                     /label=AAV2 ITR
     promoter        2878..3153
                     /label=chicken beta-actin promoter
     primer_bind     3432..3454
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3516..3535
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4358..4376)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4626..4645
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5728..5781
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5797..6495
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    6519..7107
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6572..6592)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6990..7001)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7150..7271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7161..7180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7215..7234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
       61 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      121 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      181 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      241 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      301 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      361 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      421 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      481 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      541 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      601 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      661 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      721 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      781 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      841 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      901 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      961 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1021 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1081 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1141 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1201 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1261 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1321 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1381 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1441 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1501 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1561 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1621 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1681 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1741 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1801 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1861 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     1921 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1981 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2041 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2101 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2161 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2221 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2281 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2341 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     2401 ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg
     2461 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     2521 gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta
     2581 acccgccatg ctacttatct acgtagccat gctctaggaa gatcgtacca ttgacgtcaa
     2641 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2701 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2761 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2821 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     2881 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     2941 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
     3001 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     3061 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     3121 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
     3181 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3241 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     3301 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     3361 ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
     3421 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     3481 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     3541 tcatcatttt ggcaaagaat tggatccgcc accatgtacc agaggatgct ccggtgcggt
     3601 gccgagctgg gatcgcccgg aggcggcagt ggtggcggcg caggggggcg cctggccctg
     3661 ctctggatag tcccgctcac cctcagcggc ctcctaggag tggcctgggg ggcatccagt
     3721 ttgggagcgc accacatcca ccatttccat ggcagcagca agcatcattc agtgcctatt
     3781 gcaatctaca ggtcaccagc atccttgcga ggcggacacg ctgggacgac gtatatcttt
     3841 agcaaaggtg gtggacaaat tacatataag tggcctccta atgaccgacc cagtacacga
     3901 gcagacaggc tggccatcgg gtttagcact gttcagaagg aagcagtgtt ggtgcgtgtg
     3961 gacagttcct cgggcctggg tgactacctt gagctgcaca tacaccaagg aaaaattgga
     4021 gttaaattta atgttgggac agatgacatc gccattgagg agtccaatgc aatcattaat
     4081 gatggaaaat accatgtagt gcgtttcacg aggagtggtg gcaatgccac gttacaggtg
     4141 gacagctggc cagtcatcga acgataccct gcatccggag gaggttcagg tggaggttca
     4201 ggaggaggaa tgggtggaac aagcatggtg agcaagagcg aggaggtcat caaagagttc
     4261 atgcgcttca aggtgcgctt ggagggctcc atgaacggcc acgagttcga gatcgagggc
     4321 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     4381 ggccccctgc ccttcgcctg ggacatcctg tccccccaga tcatgtacgg ctccaaggcg
     4441 tacgtgaagc accccgccga cgtccccgat tacatgaagc tgtccttccc cgagggcttc
     4501 aagtgggagc gcgtgatgca cttcgaggac ggcggtctgg tgaccgtgac tcaggacacc
     4561 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     4621 gacggccccg taatgcagaa gaagaccttg ggctgggatt atgccaccga gcgcctgtac
     4681 cccgaagacg gcgtgctgaa gggcgagctt ctggggcgcc tgaagctgaa ggacggcggc
     4741 ctcaacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     4801 tactacttcg tggacaccaa gctgggcatc acctcccaca acgaggacta caccatcgtg
     4861 gaacagtacg agcgctccga gggccgccac cacctggggc atggcaccgg cagcaccggc
     4921 agcggcagct ccggtaccgc ctcctccgag gacaacaacg gtggaggttc tggaggtggt
     4981 tctggaggtt ccggagggcg tcagctcaca atcttcaata gccaagcaac cataataatt
     5041 ggcgggaaag agcagggcca gcccttccag ggccagctct ctgggcttta ctacaatggc
     5101 ttgaaagttc tgaatatggc ggcagagaac gatgccaaca tcgccatagt gggaaatgtg
     5161 aggctggtcg gtgaagtgcc ttcctctatg acaactgagt cgacagccac tgccatgcag
     5221 tccgagatgt ccacctcaat catggagacc accacaaccc tggctaccag cacagctcga
     5281 cgaggaaagc cccccacaaa ggagcctatc agccagacca cggatgatat ccttgtggcc
     5341 tcggcagagt gtcccagcga cgatgaggac attgacccct gtgagccgag ctcagccaac
     5401 cccacccgag tgggcggccg cgaaccatac ccaggctcgg cagaggtgat ccgggagtct
     5461 agcagtacca ctggcatggt ggtggggatt gtcgcagcag cagctctgtg catcctcatc
     5521 ctcctctatg ccatgtacaa gtacagaaac cgggatgaag gttcatatca cgtggatgag
     5581 agtcgaaact acatcagtaa ctcagcacag tccaatgggg ctgtggtcaa ggagaagcag
     5641 cccagcagtg cgaaaagcgc caacaaaaac aagaagaaca aggataagga gtattatgtc
     5701 agatctggtg gcagcggtgg cagatctgag ggtcggggct ctctgctcac atgtggcgac
     5761 gtcgaggaga atcccggacc ggccccgggg tcgacaatga gcgagctgat taaggagaac
     5821 atgcacatga agctatacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc
     5881 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     5941 ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc
     6001 ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc
     6061 acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
     6121 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc
     6181 aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac
     6241 cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc
     6301 catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag
     6361 atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag
     6421 acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg
     6481 gggcacaagc taaatctgta caagtaaagc ttatcgataa tcaacctctg gattacaaaa
     6541 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     6601 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     6661 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6721 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     6781 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6841 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6901 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6961 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     7021 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     7081 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag
     7141 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     7201 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     7261 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     7321 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc
     7381 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     7441 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga c
//
