LOCUS       sequence-478959-        7067 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses green #2.4 pre-SynapShot and TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7067)
  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 7067)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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: #2.4 pre-G-SynapShot-TagBFP
FEATURES             Location/Qualifiers
     source          1..7067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        443..719
                     /label=chicken beta-actin promoter
     primer_bind     998..1020
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1082..1101
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1360..1378)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1628..1647
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1822..1845
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2811..3509
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    3533..4121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3586..3606)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4004..4015)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4164..4285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4229..4248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4791..5379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4871..4890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5550..6341,6342..6410))
                     /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     6173..6192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6411..6515)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6541..6996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6678..6699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6890..6909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg taccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      481 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      541 atgggggcgg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tccgccacca
     1141 tgccaccttc tactagtctt ttgttgcttg ctgcacttct ccccttcgct cttcccgctt
     1201 cagattggaa gacaggagaa gtcaccatgg tgagcaagag cgaggaggtc atcaaagagt
     1261 tcatgcgctt caaggtgcgc ttggagggct ccatgaacgg ccacgagttc gagatcgagg
     1321 gcgagggcga gggccgccct tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1381 gcggccccct gcccttcgcc tgggacatcc tgtcccccct gatcatgtac ggctccaaga
     1441 tgtacgtgaa gcaccccgcc gacgtccccg attacatgaa gctgtccttc cccgagggct
     1501 tcaagtggga gcgcgtgatg cacttcgagg acggcggtct ggtgaccgcg acacaggact
     1561 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     1621 ccgacggccc cgtaatgcag aagaagacct tgggctggga ttatgccacc gagcgcctgt
     1681 accccgaaga aggcgtgctg aagggcgagc tcctggggcg cctgaagctg aaggacggcg
     1741 gcctcaacct ggtggagtcc aagaccatct acatggccaa gaagcccgtg caactgcccg
     1801 gctactactt cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     1861 tggaacagta cgagcgctcc gagggccgcc accacctggg gcatggtaca ggtagcacag
     1921 gcagcggtag ctccggtacc ctgtacaagg gcggtggcgg atctggcggc ggtggtagcg
     1981 tcgactttca gaaagctagc tctattgttt ataagaagga gggggaacag gtggagttca
     2041 gtttcccatt ggccttcaca gtcgagaagc tcacaggatc aggagagttg tggtggcagg
     2101 ccgaaagagc ctcctccagc aagagttgga tcacttttga tctgaaaaac aaagaagtga
     2161 gcgtgaagag agtcacacag gaccctaagc tgcagatggg caagaagttg cctttgcact
     2221 tgacactccc ccaagccctg ccccagtatg ctggctccgg gaacctcacc ctcgccttgg
     2281 aggctaagac cggaaagttg catcaggagg tcaacctcgt ggtgatgagg gcaacacagc
     2341 tgcagaagaa cctgacctgc gaagtgtggg gtcccacctc acctaaactg atgctgtctc
     2401 tgaagctgga gaataaggag gctaaagtca gtaaaagaga gaaggctgtg tgggtcctta
     2461 acccggaggc agggatgtgg cagtgtcttc ttagcgactc tggacaggtg ctgctggaga
     2521 gcaacatcaa ggtgcttcct acttggtcaa caccagtaca gccgatggcc ctgatcgtac
     2581 tcgggggagt cgccggcctg cttctgttta tcggcctggg catcttcttc tgcgtacgct
     2641 acaagtatcg caacagggat gagggatctt accacgttga cgaatctagg aactatatca
     2701 gtaacagcgc tcagagtaat ggcgccgtgg ttaaagagaa acagccctcc agcgccaaat
     2761 cagcaaacaa aaacaagaaa aacaaggata aagagtacta cgtaagatct atgagcgagc
     2821 tgattaagga gaacatgcac atgaagctat acatggaggg caccgtggac aaccatcact
     2881 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     2941 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct
     3001 acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct
     3061 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     3121 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     3181 tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca
     3241 ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc
     3301 tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg
     3361 ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg
     3421 aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc
     3481 tccctagcaa actggggcac aagctaaatc tgtacaagta aagcttatcg ataatcaacc
     3541 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3601 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3661 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3721 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3781 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3841 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3901 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3961 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4021 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4081 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg
     4141 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4201 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4261 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4321 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4381 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     4441 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4501 ctgaggccgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     4561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4981 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5221 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5461 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6541 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6601 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     6661 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     6721 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     6781 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6841 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6901 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6961 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     7021 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccag
//
