LOCUS       Exported                7339 bp ds-DNA     circular SYN 06-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 10, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7339
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          5001..5042
     misc_feature    complement(23..187)
                     /label=ITR
                     /note="ITR"
     misc_feature    complement(194..236)
                     /label=seq
                     /note="seq"
     misc_feature    complement(251..270)
                     /label=pzac2.1 5088f
                     /note="pzac2.1 5088f"
     primer_bind     complement(351..374)
                     /label=f.ITR for 2.01.14
                     /note="f.ITR for 2.01.14"
     primer_bind     complement(452..472)
                     /label=f.ITR for 1.01.13
                     /note="f.ITR for 1.01.13"
     primer_bind     2471..2489
                     /label=r.ITR rev 3.01.19
                     /note="r.ITR rev 3.01.19"
     primer_bind     2513..2536
                     /label=r.ITR rev 4.01.20
                     /note="r.ITR rev 4.01.20"
     misc_feature    complement(2707..2871)
                     /label=ITR
                     /note="ITR"
     promoter        2898..3832
                     /label=CAG
                     /note="CAG "
     primer_bind     2975..2994
                     /label=f.CAG promoter seq.33.1743
                     /note="f.CAG promoter seq.33.1743"
     primer_bind     complement(3043..3058)
                     /label=r.CAG-flex ITR seq.33.1306 CAG rev
                     /note="r.CAG-flex ITR seq.33.1306 CAG rev"
     primer_bind     3049..3066
                     /label=f.CAG promoter.33.1744
                     /note="f.CAG promoter.33.1744"
     primer_bind     3483..3499
                     /label=f.CAG promoter seq.33.1747
                     /note="f.CAG promoter seq.33.1747"
     primer_bind     3495..3516
                     /label=r.CAG promoter seq.33.1745
                     /note="r.CAG promoter seq.33.1745"
     primer_bind     complement(3581..3603)
                     /label=r.CAG promoter seq.33.1746
                     /note="r.CAG promoter seq.33.1746"
     primer_bind     3703..3725
                     /label=f.CAG for.01.40
                     /note="f.CAG for.01.40"
     primer_bind     complement(3795..3818)
                     /label=r.CAG promoter seq.33.1748
                     /note="r.CAG promoter seq.33.1748"
     misc_feature    3833
                     /label=pCAG_GCaMP3
     misc_feature    3840..3845
                     /label=Kozak
     CDS             3846..3857
                     /codon_start=1
                     /label=MCS
                     /note="MCS"
                     /translation="MGRS"
     CDS             3858..4184
                     /codon_start=1
                     /label=PS3 epsilon
                     /note="PS3 epsilon"
                     /translation="MKTIHVSVVTPDGPVYEDDVEMVSVKAKSGELGILPGHIPLKAPL
                     EISAARLKKGGKTQYIAVSGGNLEVRPDKVTIYAQAAERAEDIDVLRAKKAKERAERRL
                     QSQVL"
     misc_feature    3882..3884
                     /label=T9V
                     /note="T9V"
     misc_feature    4035..4037
                     /label=Y60Y
                     /note="Y60Y"
     misc_feature    4089..4091
                     /label=N78Y
                     /note="N78Y"
     misc_feature    4140..4142
                     /label=A95K
     misc_feature    4179..4184
                     /label=QD>VL
                     /note="QD>VL"
     CDS             4185..4460
                     /codon_start=1
                     /label=cpsfGFP
                     /note="cpsfGFP"
                     /translation="VGNVYITADKQKNGIKANFKIRHNVEDGSMQLADHYQQNTPIGDG
                     PVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    4185..4190
                     /label=SG>VG
                     /note="SH>VG"
     misc_feature    4272..4274
                     /label=V>M
                     /note="V->M"
     CDS             4461..4478
                     /codon_start=1
                     /label=cpLinker
                     /note="cpsfGFP"
                     /translation="GGTGGS"
     CDS             4479..4913
                     /codon_start=1
                     /label=cpSFGFP
                     /note="cpSFGFP"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNF"
     CDS             4914..4922
                     /codon_start=1
                     /label=L2-ICV
                     /note="L2+++"
                     /translation="ICV"
     CDS             4923..4994
                     /codon_start=1
                     /label=PS3 epsilon
                     /note="PS3 epsilon"
                     /translation="DIDFKRAELALKRAMNRLSVAEMK"
     misc_feature    4950..4952
                     /label=A119
     CDS             4995..5000
                     /codon_start=1
                     /label=MCS
                     /translation="LQ"
     CDS             5001..5042
                     /codon_start=1
                     /label=3xGGGGS
                     /note="/__SeqFeature__=True"
                     /note="/__level__=0"
                     /note="/application_notes=Commonly used flexible linker for
                     fusing two proteins."
                     /note="/full_name=3 tandem GGGGS"
                     /note="/name=3xGGGGS"
                     /note="/official_designation=3xGGGGS"
                     /note="/uuid=82e342ab8d8e4d71a1f1c7026d54944d"
                     /note="/vntifkey=21"
                     /translation="GGGSGGGGSGGGGS"
     CDS             5043..5720
                     /codon_start=1
                     /label=tdTomato Part1
                     /note="tdTomato Part1"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             5721..5777
                     /codon_start=1
                     /label=Linker
                     /note="Linker"
                     /translation="GHGTGSTGSGSSGTASSED"
     CDS             5778..6470
                     /codon_start=1
                     /label=tdTomato part2
                     /note="tdTomato part2"
                     /translation="NNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAKLK
                     VTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGLVT
                     VTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQAL
                     KLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHLF
                     LYGMDELYK"
     misc_feature    6486..7074
                     /label=WPRE
                     /note="WPRE"
     primer_bind     complement(6557..6576)
                     /label=r.wpre rev.01.3
                     /note="r.wpre rev.01.3"
     primer_bind     complement(6887..6907)
                     /label=wpre rev pcr 32.63
                     /note="wpre rev pcr 32.63"
     primer_bind     6911..6930
                     /label=f.WPRE seq.01.80
                     /note="f.WPRE seq.01.80"
     primer_bind     7118..7141
                     /label=f.ITR for 3.01.15
                     /note="f.ITR for 3.01.15"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2341 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2401 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2461 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2521 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2581 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tcgtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2941 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3001 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3061 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3121 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     3181 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     3241 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     3301 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     3361 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     3421 gcgggagtcg ctgcgacgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     3481 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     3541 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     3601 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc
     3661 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     3721 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     3781 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttggatcccg
     3841 ccaccatggg cagatctatg aagactattc acgtgagtgt cgtaactccc gacgggcctg
     3901 tatatgagga tgacgttgaa atggtgagcg tcaaagcaaa aagtggcgag ctcggtattc
     3961 tcccaggcca cattcccttg aaagctcccc tagagatcag tgccgcacgc ctgaagaaag
     4021 ggggcaaaac acagtatatc gctgtgtcag gcggcaacct cgaagtgcgg cctgacaagg
     4081 tgaccatcta tgcccaggca gccgagaggg ctgaggatat cgatgtcctg cgcgccaaga
     4141 aagccaagga gagagccgag cgccgactcc aatcacaggt cctggtcggg aacgtctata
     4201 tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacgtgg
     4261 aggacggcag catgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4321 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgtgctgagc aaagacccta
     4381 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4441 gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc gaggagctgt
     4501 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     4561 gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     4621 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     4681 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     4741 tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc acctacaaga
     4801 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     4861 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tttatttgtg
     4921 tggacatcga ttttaagcgc gccgagctcg cccttaagcg cgcaatgaat aggctctcag
     4981 ttgccgaaat gaagctgcag ggcggaggct cgggcggagg tgggtcgggt ggcggcggat
     5041 caatggttag caagggtgaa gaggtgatta aagagtttat gcgttttaaa gtacgcatgg
     5101 agggtagcat gaacggtcat gagtttgaga tcgagggcga aggcgaaggt cgtccgtacg
     5161 agggtacgca gaccgctaaa ctgaaagtaa ccaaaggcgg tcctctgccg ttcgcgtggg
     5221 acatcctgtc cccgcagttt atgtatggtt ccaaggccta cgtcaaacac ccagccgata
     5281 tcccggacta caagaagctg tctttcccgg agggtttcaa gtgggagcgt gtaatgaact
     5341 ttgaggacgg tggcctggtt accgtcaccc aggatagctc tctgcaagat ggcaccctga
     5401 tctacaaggt gaagatgcgt ggtacgaact ttccgccgga tggcccggtg atgcagaaaa
     5461 agacgatggg atgggaagca tctaccgagc gcctgtatcc gcgtgacggt gttctgaaag
     5521 gtgagatcca ccaggcgctg aaactgaagg acggcggtca ctacctggtt gaatttaaga
     5581 cgatctatat ggccaaaaag ccggttcaac tgccaggtta ctactacgtg gacaccaaac
     5641 tggatatcac gtctcacaac gaagattata ccatcgtcga acagtatgaa cgtagcgagg
     5701 gccgtcacca cctgttcctg ggccacggta ctggttctac gggttctggt tcctctggca
     5761 ctgcgtcttc cgaagacaac aacatggcgg tgatcaaaga gttcatgagg ttcaaagtac
     5821 gtatggaagg ttctatgaac ggccatgagt tcgagattga aggtgagggc gagggtcgcc
     5881 catacgaggg cactcagacc gcaaagctga aagttaccaa aggtggccca ctgccttttg
     5941 cgtgggatat cctgtctccg cagttcatgt acggttccaa agcctacgta aaacacccgg
     6001 cagacatccc ggattacaaa aagctgtcct tcccagaggg cttcaaatgg gaacgtgtta
     6061 tgaatttcga agatggcggc ctggtcactg ttacgcagga cagcagcctg caagacggta
     6121 ctctgattta caaagtgaaa atgcgtggca ctaacttccc gcctgacggt ccagtaatgc
     6181 agaagaaaac tatgggctgg gaagctagca ccgaacgtct gtacccgcgc gacggtgtgc
     6241 tgaaaggcga gattcaccag gccctgaagc tgaaggatgg tggtcactat ctggttgagt
     6301 tcaaaactat ctacatggca aaaaagcctg tacaactgcc gggctattac tacgttgaca
     6361 ctaaactgga catcacctcc cacaacgagg attatactat tgttgaacag tacgagcgtt
     6421 ctgaaggtcg ccatcatctg tttctgtatg gtatggacga actgtataaa tagaagctta
     6481 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     6541 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     6601 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     6661 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     6721 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6781 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6841 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     6901 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6961 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     7021 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     7081 accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     7141 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     7201 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     7261 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     7321 aatgtggtaa aatcgataa
//
