LOCUS       sequence-311050-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes pAAV NovArch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  AUTHORS   Chien MP, Brinks D, Testa-Silva G, Tian H, Phil Brooks F 3rd, Adam 
            Y, Bloxham W, Gmeiner B, Kheifets S, Cohen AE
  TITLE     Photoactivated voltage imaging in tissue with an 
            archaerhodopsin-derived reporter.
  JOURNAL   Sci Adv. 2021 May 5;7(19). pii: 7/19/eabe3216. doi: 
            10.1126/sciadv.abe3216. Print 2021 May.
  PUBMED    33952514
REFERENCE   2  (bases 1 to 6816)
  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: "Sci Adv."; 
            date: "2021-05-5"; volume: "7(19). pii: 7/19/eabe3216. doi"; pages: 
            " 10.1126/sciadv.abe3216. Print 2021 May"
COMMENT     Sequence Label: MPC026: pAAV-CAG-FLEX-NovArch-TS-Citrine_TSx3-ER2
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..718
                     /label=chicken beta-actin promoter
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1135..1168
                     /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    1231..1264
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1517..2227,2228..2230))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1559..1580)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1908..1927
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2167..2188
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2300..3058)
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILASGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAARERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVIGLGTETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     regulatory      3055..3064
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3081..3114)
                     /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(3185..3218)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3260..3848
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3313..3333)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3731..3742)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3891..4012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3902..3921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3956..3975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4174..4303
                     /label=AAV2 ITR
     rep_origin      complement(4540..5128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4620..4639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5299..6090,6091..6159))
                     /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     5922..5941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6160..6264)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6290..6745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6427..6448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6639..6658
                     /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 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg gggggggggg
      601 gggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1261 ttatactagg gatcccccct cgagttttac ttgtacatta cacttcattc tcataacaaa
     1321 agccccccac attgatgtca atctgatcga ggggtatata ctcgccctca gaagtgattc
     1381 gactttttcc ccccacgttt atatcgattt gatccagagg gatgtactcg ccctcagagg
     1441 ttatgcggga ttttccaccg acgttaatgt cgatctgatc cagagggata tactcgccct
     1501 cagaggtgat ccttgatttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     1561 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     1621 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     1681 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     1741 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     1801 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1861 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     1921 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     1981 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcacatcagg ccgtagccga
     2041 aggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     2101 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     2161 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     2221 ccttgctcac tacatttatg tctatttgat ccagggggat gtactcgcct tcgcttgtga
     2281 ttctactctt catcaacgta tcggcaccgg cactgggttc tggtgcctcg gtgtcgccca
     2341 gaatagcccg ggatctcaac aggataaagc caaagccgac cttggcagtc acgtccaaca
     2401 ccataaacag cagagtttcg gtgcccaggc ctatcacgcc agcgccctca gtgcctatga
     2461 tccacaggat agggtaagcg gtccacagca ccaagaccag agctgtcagg gtgttaaagg
     2521 tagatgccac ctcggggccc cgctcccttg cagcagatcg cagggatgta gccagaacat
     2581 agagcaccac tatcatgcaa attgtagaga acaaccacca actgtatctg gctatggccg
     2641 tgtggctcaa ggctccgatg aggccagtga cgatcatcag ggcgtccaca cccaccaggg
     2701 tgccgatggt cacccgatcc accttagcga gaagggccag gtgcagcagc agaagtgggg
     2761 tggtaaacag ccactgggcg tacctggcat aatagatatc caacatttcg cccccgacgg
     2821 acacctcagt aagcccgata ccaaagaaca tagacagata tgcggcggat gcgattccgg
     2881 aggccaggat agtcacagcg taatattccc gggcatcctt atcggtgact ccccatccgc
     2941 ggaccagaaa gtagaaggtt ccaatcagca tcagcagagt gccgatgccc agccacaggg
     3001 tttcaggtct gccgtcaccc agtaggtcgt aaccagcctg cagagcgata cttaccatgg
     3061 tggcggtacc gaattctagt ataacttcgt ataggatact ttatacgaag ttatcattgg
     3121 gattcttcct attttgatcc aagcatcacc atcgaccctc tagtccagat ctcaccatcg
     3181 acccataact tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgaatt
     3241 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3301 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3361 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3421 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3481 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3541 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3601 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3661 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3721 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3781 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3841 ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat
     3901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4021 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     4081 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     4141 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     4201 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4261 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     6301 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     6361 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     6421 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6481 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6541 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6601 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6661 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6721 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     6781 gggcgatcgg tgcgggcctc ttcgctatta cgccag
//
