LOCUS       sequence-426470-        7557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses green #1.4 pre-SynapShot and TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7557)
  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 7557)
  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-G-SynapShot-TagBFP
FEATURES             Location/Qualifiers
     source          1..7557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   32..161
                     /label=AAV2 ITR
     promoter        474..751
                     /label=chicken beta-actin promoter
     primer_bind     1030..1052
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1114..1133
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1953..1971)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2221..2240
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2415..2438
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3263..3316
                     /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             3332..4030
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    4054..4642
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4107..4127)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4525..4536)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4685..4806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4696..4715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4750..4769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5147..5602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5234..5253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5444..5465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5628..5732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5733..5801,5802..6593)
                     /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     complement(5951..5970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6764..7352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7253..7272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctgcgcgct cgctcgctca ctgaggccgc
       61 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
      121 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
      181 gccatgctac ttatctacgt agccatgctc taggaagatc gtaccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
      481 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
      541 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
      601 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
      661 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
      721 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
      781 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
      841 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
      901 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
      961 gtgcgggggg agcggctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
     1021 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
     1081 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1141 atcattttgg caaagaattg gatccgccac catgtaccag aggatgctcc ggtgcggtgc
     1201 cgagctggga tcgcccggag gcggcagtgg tggcggcgca ggggggcgcc tggccctgct
     1261 ctggatagtc ccgctcaccc tcagcggcct cctaggagtg gcctgggggg catccagttt
     1321 gggagcgcac cacatccacc atttccatgg cagcagcaag catcattcag tgcctattgc
     1381 aatctacagg tcaccagcat ccttgcgagg cggacacgct gggacgacgt atatctttag
     1441 caaaggtggt ggacaaatta catataagtg gcctcctaat gaccgaccca gtacacgagc
     1501 agacaggctg gccatcgggt ttagcactgt tcagaaggaa gcagtgttgg tgcgtgtgga
     1561 cagttcctcg ggcctgggtg actaccttga gctgcacata caccaaggaa aaattggagt
     1621 taaatttaat gttgggacag atgacatcgc cattgaggag tccaatgcaa tcattaatga
     1681 tggaaaatac catgtagtgc gtttcacgag gagtggtggc aatgccacgt tacaggtgga
     1741 cagctggcca gtcatcgaac gataccctgc atccggagga ggttcaggtg gaggttcagg
     1801 aggaggaatg ggtggaacaa gcgtgagcaa gagcgaggag gtcatcaaag agttcatgcg
     1861 cttcaaggtg cgcttggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg
     1921 cgagggccgc ccttacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc
     1981 cctgcccttc gcctgggaca tcctgtcccc cctgatcatg tacggctcca agatgtacgt
     2041 gaagcacccc gccgacgtcc ccgattacat gaagctgtcc ttccccgagg gcttcaagtg
     2101 ggagcgcgtg atgcacttcg aggacggcgg tctggtgacc gcgacacagg actcctccct
     2161 gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg
     2221 ccccgtaatg cagaagaaga ccttgggctg ggattatgcc accgagcgcc tgtaccccga
     2281 agaaggcgtg ctgaagggcg agctcctggg gcgcctgaag ctgaaggacg gcggcctcaa
     2341 cctggtggag tccaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta
     2401 cttcgtggac accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2461 gtacgagcgc tccgagggcc gccaccacct ggggcatggt acaggtagca caggcagcgg
     2521 tagctccggt gggcgtcagc tcacaatctt caatagccaa gcaaccataa taattggcgg
     2581 gaaagagcag ggccagccct tccagggcca gctctctggg ctttactaca atggcttgaa
     2641 agttctgaat atggcggcag agaacgatgc caacatcgcc atagtgggaa atgtgaggct
     2701 ggtcggtgaa gtgccttcct ctatgacaac tgagtcgaca gccactgcca tgcagtccga
     2761 gatgtccacc tcaatcatgg agaccaccac aaccctggct accagcacag ctcgacgagg
     2821 aaagcccccc acaaaggagc ctatcagcca gaccacggat gatatccttg tggcctcggc
     2881 agagtgtccc agcgacgatg aggacattga cccctgtgag ccgagctcag ccaaccccac
     2941 ccgagtgggc ggccgcgaac catacccagg ctcggcagag gtgatccggg agtctagcag
     3001 taccactggc atggtggtgg ggattgtcgc agcagcagct ctgtgcatcc tcatcctcct
     3061 ctatgccatg tacaagtaca gaaaccggga tgaaggttca tatcacgtgg atgagagtcg
     3121 aaactacatc agtaactcag cacagtccaa tggggctgtg gtcaaggaga agcagcccag
     3181 cagtgcgaaa agcgccaaca aaaacaagaa gaacaaggat aaggagtatt atgtcagatc
     3241 tggtggcagc ggtggcagat ctgagggtcg gggctctctg ctcacatgtg gcgacgtcga
     3301 ggagaatccc ggaccggccc cggggtcgac aatgagcgag ctgattaagg agaacatgca
     3361 catgaagcta tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg
     3421 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
     3481 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat
     3541 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg
     3601 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     3661 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg
     3721 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc
     3781 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct
     3841 gatcgcaaac atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc
     3901 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta
     3961 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca
     4021 caagctaaat ctgtacaagt aaagcttatc gataatcaac ctctggatta caaaatttgt
     4081 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4141 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4201 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4261 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4321 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4381 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4441 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     4501 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttccctcggc
     4561 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4621 tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac
     4681 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     4741 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     4801 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     4861 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga
     4921 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     4981 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgasgmcg cccgggcttt
     5041 gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc gtaatagcga agaggcccgc
     5101 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     5161 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5221 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5281 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5341 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5401 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5461 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5521 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5581 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6901 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7381 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7441 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7501 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctc
//
