LOCUS       sequence-428987-        7068 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 7068)
  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 7068)
  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..7068
                     /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(1392..1410)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1660..1679
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1854..1877
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2843..3541
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    3565..4153
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3618..3638)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4036..4047)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4196..4317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4207..4226)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4261..4280
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4658..5113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4745..4764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4955..4976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5139..5243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5244..5312,5313..6104)
                     /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(5462..5481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6275..6863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6764..6783
                     /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 catgccacct tctactagtc ttttgttgct
     1201 tgctgcactt ctccccttcg ctcttcccgc ttcagattgg aagacaggag aagtcaccat
     1261 ggtgagcaag agcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcttggaggg
     1321 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cttacgaggg
     1381 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     1441 cctgtccccc ctgatcatgt acggctccaa gatgtacgtg aagcaccccg ccgacgtccc
     1501 cgattacatg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgcacttcga
     1561 ggacggcggt ctggtgaccg cgacacagga ctcctccctg caggacggca cgctgatcta
     1621 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     1681 cttgggctgg gattatgcca ccgagcgcct gtaccccgaa gaaggcgtgc tgaagggcga
     1741 gctcctgggg cgcctgaagc tgaaggacgg cggcctcaac ctggtggagt ccaagaccat
     1801 ctacatggcc aagaagcccg tgcaactgcc cggctactac ttcgtggaca ccaagctgga
     1861 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     1921 ccaccacctg gggcatggta caggtagcac aggcagcggt agctccggta ccctgtacaa
     1981 gggcggtggc ggatctggcg gcggtggtag cgtcgacttt cagaaagcta gctctattgt
     2041 ttataagaag gagggggaac aggtggagtt cagtttccca ttggccttca cagtcgagaa
     2101 gctcacagga tcaggagagt tgtggtggca ggccgaaaga gcctcctcca gcaagagttg
     2161 gatcactttt gatctgaaaa acaaagaagt gagcgtgaag agagtcacac aggaccctaa
     2221 gctgcagatg ggcaagaagt tgcctttgca cttgacactc ccccaagccc tgccccagta
     2281 tgctggctcc gggaacctca ccctcgcctt ggaggctaag accggaaagt tgcatcagga
     2341 ggtcaacctc gtggtgatga gggcaacaca gctgcagaag aacctgacct gcgaagtgtg
     2401 gggtcccacc tcacctaaac tgatgctgtc tctgaagctg gagaataagg aggctaaagt
     2461 cagtaaaaga gagaaggctg tgtgggtcct taacccggag gcagggatgt ggcagtgtct
     2521 tcttagcgac tctggacagg tgctgctgga gagcaacatc aaggtgcttc ctacttggtc
     2581 aacaccagta cagccgatgg ccctgatcgt actcggggga gtcgccggcc tgcttctgtt
     2641 tatcggcctg ggcatcttct tctgcgtacg ctacaagtat cgcaacaggg atgagggatc
     2701 ttaccacgtt gacgaatcta ggaactatat cagtaacagc gctcagagta atggcgccgt
     2761 ggttaaagag aaacagccct ccagcgccaa atcagcaaac aaaaacaaga aaaacaagga
     2821 taaagagtac tacgtaagat ctatgagcga gctgattaag gagaacatgc acatgaagct
     2881 atacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa
     2941 gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt
     3001 cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac
     3061 ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt
     3121 caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg
     3181 ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat
     3241 gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg
     3301 cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa
     3361 catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta
     3421 ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct acgtcgagca
     3481 gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc acaagctaaa
     3541 tctgtacaag taaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3601 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3661 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3721 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3781 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3841 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3901 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3961 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4021 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4081 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4141 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga
     4201 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4261 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4321 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     4381 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     4441 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4501 ggccactccc tctctgcgcg ctcgctcgct cacggccgcc gcccgggctt tgcccgggcg
     4561 gcctcagtga gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc
     4621 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4681 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4741 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4801 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4861 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4921 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4981 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5041 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5101 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5221 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5281 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5341 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5401 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5461 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5521 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5581 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5641 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5701 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5761 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5821 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5881 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5941 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6001 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6061 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6121 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6181 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6241 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6301 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6361 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6421 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6721 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6781 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6841 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6901 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6961 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7021 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctc
//
