LOCUS       Exported                6351 bp ds-DNA     circular SYN 04-DEC-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 13, 2025 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6351
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          4988..5029
     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"
     primer_bind     3833..3869
                     /label=ATPsnifer2 BamHI Kozak HiFi F
     misc_feature    3833
                     /label=pCAG_GCaMP3
     misc_feature    3839..3844
                     /label=Kozak
     CDS             3845..4171
                     /codon_start=1
                     /label=PS3 epsilon
                     /note="PS3 epsilon"
                     /translation="MKTIHVSVVTPDGPVYEDDVEMVSVKAKSGELGILPGHIPLKAPL
                     EISAARLKKGGKTQYIAVSGGNLEVRPDKVTIYAQAAERAEDIDVLRAKAAKERAERRL
                     QSQVL"
     misc_feature    3869..3871
                     /label=T9V
                     /note="T9V"
     misc_feature    4022..4024
                     /label=Y60Y
                     /note="Y60Y"
     misc_feature    4076..4078
                     /label=N78Y
                     /note="N78Y"
     misc_feature    4127..4129
                     /label=A95K
     misc_feature    4166..4171
                     /label=QD>VL
                     /note="QD>VL"
     CDS             4172..4447
                     /codon_start=1
                     /label=cpsfGFP
                     /note="cpsfGFP"
                     /translation="VGNVYITADKQKNGIKANFKIRHNVEDGSMQLADHYQQNTPIGDG
                     PVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    4172..4177
                     /label=SG>VG
                     /note="SH>VG"
     misc_feature    4259..4261
                     /label=V>M
                     /note="V->M"
     CDS             4448..4465
                     /codon_start=1
                     /label=cpLinker
                     /note="cpsfGFP"
                     /translation="GGTGGS"
     CDS             4466..4900
                     /codon_start=1
                     /label=cpSFGFP
                     /note="cpSFGFP"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNF"
     primer_bind     4516..4526
                     /label=miRFP670nano3 Stop EcoRI HiFi R
     CDS             4901..4909
                     /codon_start=1
                     /label=L2-ICV
                     /note="L2+++"
                     /translation="ICV"
     CDS             4910..4981
                     /codon_start=1
                     /label=PS3 epsilon
                     /note="PS3 epsilon"
                     /translation="DIDFKRAELLLKRAMNRLSVAEMK"
     misc_feature    4937..4939
                     /label=A119L
     CDS             4982..4987
                     /codon_start=1
                     /label=MCS
                     /translation="LQ"
     CDS             4988..5029
                     /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"
     primer_bind     complement(5014..5042)
                     /label=ATPsnifer2 HiFi R
     primer_bind     5018..5050
                     /label=miRFP670nano3 HiFi F
     CDS             5030..5470
                     /codon_start=1
                     /label=miRFP670nano3
                     /translation="MANLDKMLNTTVTEVRKFLQADRVCVFKFEEDYSGTVSHEAVDDR
                     WISILKTQVQDRYFMETRGEEYVHGRYQAIADIYTANLVECYRDLLIEFQVRAILAVPI
                     LQGKKLWGLLVAHQLAGPREWQTWEIDFLKQQAVVMGIAIQQS"
     primer_bind     complement(5455..5503)
                     /label=miRFP670nano3 Stop EcoRI HiFi R
     misc_feature    5498..6086
                     /label=WPRE
                     /note="WPRE"
     primer_bind     complement(5569..5588)
                     /label=r.wpre rev.01.3
                     /note="r.wpre rev.01.3"
     primer_bind     complement(5899..5919)
                     /label=wpre rev pcr 32.63
                     /note="wpre rev pcr 32.63"
     primer_bind     5923..5942
                     /label=f.WPRE seq.01.80
                     /note="f.WPRE seq.01.80"
     primer_bind     6130..6153
                     /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 ttggatccgc
     3841 caccatgaag actattcacg tgagtgtcgt aactcccgac gggcctgtat atgaggatga
     3901 cgttgaaatg gtgagcgtca aagcaaaaag tggcgagctc ggtattctcc caggccacat
     3961 tcccttgaaa gctcccctag agatcagtgc cgcacgcctg aagaaagggg gcaaaacaca
     4021 gtatatcgct gtgtcaggcg gcaacctcga agtgcggcct gacaaggtga ccatctatgc
     4081 ccaggcagcc gagagggctg aggatatcga tgtcctgcgc gccaaggccg ccaaggagag
     4141 agccgagcgc cgactccaat cacaggtcct ggtcgggaac gtctatatca ccgccgacaa
     4201 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacgtggagg acggcagcat
     4261 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4321 cgacaaccac tacctgagca cccagtccgt gctgagcaaa gaccctaacg agaagcgcga
     4381 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4441 gtacaagggc ggtaccggag ggagcatgag caagggcgag gagctgttca ccggggtggt
     4501 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgcgcggcga
     4561 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4621 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4681 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4741 cgtccaggag cgcaccatca gcttcaagga cgacggcacc tacaagaccc gcgccgaggt
     4801 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4861 ggacggcaac atcctggggc acaagctgga gtacaacttt atttgtgtgg acatcgattt
     4921 taagcgcgcc gagctcttgc ttaagcgcgc aatgaatagg ctctcagttg ccgaaatgaa
     4981 gctgcagggc ggaggctcgg gcggaggtgg gtcgggtggc ggcggatcaa tggcaaacct
     5041 ggacaagatg ctgaacacca ccgtgaccga ggtgcgcaag ttcctgcaag cggacagagt
     5101 gtgcgtgttc aagttcgagg aagattactc cggcaccgtc tcgcacgaag ccgtggacga
     5161 cagatggatt agcatcctga agacccaggt gcaggacaga tacttcatgg aaaccagagg
     5221 cgaggaatac gtccacggca gataccaggc catcgccgac atctacacag ccaatctggt
     5281 cgagtgctac agagacctgc tgatcgagtt tcaggtgcgg gccattctgg ctgtccccat
     5341 cctgcaaggc aagaagctgt ggggcctgct ggtggcccac caactggccg gccctcggga
     5401 gtggcagacc tgggaaatcg acttcctgaa acagcaagcc gtggtgatgg gcatcgccat
     5461 ccagcagagc taggaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     5521 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5581 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5641 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5701 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5761 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5821 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5881 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5941 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6001 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6061 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     6121 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     6181 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     6241 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     6301 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata a
//
