LOCUS       sequence-459097-        7297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses green #2.4 post-SynapShot and mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7297)
  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 7297)
  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: #2.4 post-G-SynapShot-mScarlet
FEATURES             Location/Qualifiers
     source          1..7297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2899..3176
                     /label=chicken beta-actin promoter
     primer_bind     3455..3477
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3539..3558
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    3593..3626
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3689..3722
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3748..4443)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     CDS             complement(4459..4515)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4888..4899
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      5994..6003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6020..6053)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(6124..6157)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6199..6787
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6252..6272)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6670..6681)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6830..6951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6841..6860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6895..6914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7113..7242
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2761 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2821 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2881 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     2941 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     3001 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
     3061 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     3121 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     3181 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     3241 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     3301 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     3361 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     3421 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3481 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3541 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc ccataacttc
     3601 gtataaagta tcctatacga agttatatca aaataggaag accaatgctt caccatcgac
     3661 ccgaattgcc aagcatcacc atcgacccat aacttcgtat aatgtatgct atacgaagtt
     3721 atactaggga tcccccctcg agttttactt gtacagctcg tccatgccgc cggtggagtg
     3781 gcggccctcg gagcgttcgt actgttccac cacggtgtag tcctcgttgt gggaggtgat
     3841 gtccaacttg cggtcgacgt tgtaggcgcc gggcatctgc acgggcttct tggccttgta
     3901 ggtggtcttg aagtccgcca ggtagcggcc gccgtccttc aggcgcaggg ccatcttaat
     3961 gtcgcccttc agcacgccgt cctcggggta caaccgctcg gtggacgctt cccagcccat
     4021 tgtcttcttc tgcattacgg ggccgtcagg agggaagttg gtgccgcgga gcttcacctt
     4081 gtagatcagg gtgccgtcct ccagggaggt gtcctgggtc acggtcacgg cgccgccgtc
     4141 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggactgct tatagtagtc
     4201 ggggatgtcg gcggggtgct tggtgaaggc cctggagccg tacatgaact gaggggacag
     4261 gatgtcccag gagaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
     4321 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga
     4381 gccctccatg tgcaccttga accgcatgaa ctccttgatc actgcctcgc ccttgctcac
     4441 cattgtcgac cccggggccg gtccagggtt ctcctccacg tctccagcct gcttcagcag
     4501 gctgaagtta gtagctccgc ttccgctgtg ggagtggggg tgtggatgcg gatgagggag
     4561 agtattattc tgtcctccag taaatgtgtt gaatgtatgt aagggctgaa tccctggtat
     4621 agtgttggga atcattgtga tggtgttagg tgtcattaaa ggaacatcat ccggtgacct
     4681 cctcatagca agagtataat ctggggggca tgctgtcctc agcaccacct catggggatg
     4741 gatggattca cactcgtggt ccaggtctgt gtgtttcatc tgaagactca tgatctcctc
     4801 ctcaggtgcg tgcgtaagat cattggttgt agttcgttgc ggactacatc tcctatgtac
     4861 gtcgtggcgc cttttgtcct tcttgtaata gagggccgcg aaagccagaa tgttcaggaa
     4921 aagcagggaa gctcctacag ctatggtcac ggacagctct gtgctgtagt ccctttggtc
     4981 tacgctaaat ggactaggct gttgtttggg gtcgtcttgc ttagcggttg gaaatgcgct
     5041 ggtcacaggg gttgagtttt tccgagttgg tcgaagggtg atgtcagtgc ttgggacctt
     5101 ggtggttgta cttgtgtatt ggctgatgtc attgaggtta tgcaagtggg gcaccagctc
     5161 gagcgtcccc ttgtaccgct cgtccatgcc cagacggtgg cggccctcgg agcgcccgta
     5221 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccagcttgg tgtccacaca
     5281 gtaatcgccg ggcagttgca cgggcttctt ggccatgtag atggtctcga actccaccag
     5341 gtagtggccg ccgtccttca gcttcagggc ctgatagcta tggcccttca gcacgccgtc
     5401 cttggggtac agcatctcgg tggaggcctc ccagcccatg gtcttcttct gcattacggg
     5461 gccgtcgggg gggaagttgg tgccgcgaat cttcaccttg cagatcagcg tgccgtcctg
     5521 cagggaggag tcctgggtaa cggtcaccag accgccgtct tcgaagttca tcacgcgctc
     5581 ccacttgaag ccctcgggga agggcagctt cttgtaatcg gggatgtcgg cggggtgcct
     5641 cacgtacgct ttggagccgt acatgatctg gggggacagg atgtcccagg caaagggcag
     5701 ggggccgccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     5761 gccctcgccc tcgatctcga actcgtggcc gttcatggag ccctccatgc gcaccttgaa
     5821 gcgcatgaac tctttgatgg tggccatgtc atccagcttc tgtgaaagaa cgtggaaagt
     5881 ctggaggagg caacccaaca ggcagagtgt aagtggggcg ccactccctc ggtgcaccac
     5941 gcaggccatc atagctcgcc atacataatt aggccacata catctaggaa gtgccatggt
     6001 ggcggtaccg aattctagta taacttcgta taggatactt tatacgaagt tatcattggg
     6061 attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga
     6121 cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc
     6181 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6241 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6301 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6361 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6421 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6481 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6541 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     6601 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6661 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6721 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6781 tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     6841 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     6901 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     6961 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     7021 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     7081 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     7141 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     7201 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     7261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgct
//
