LOCUS       sequence-256058-        6267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of ChRmine-mScarlet under the 
            control of human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja 
            C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, 
            Yoshizawa S, Kato HE, Ganguli S, Deisseroth K
  TITLE     Cortical layer-specific critical dynamics triggering perception.
  JOURNAL   Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 
            10.1126/science.aaw5202. Epub 2019 Jul 18.
  PUBMED    31320556
REFERENCE   2  (bases 1 to 6267)
  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: "Science."; 
            date: "2019-08-9"; volume: "365(6453). pii: science.aaw5202. doi"; 
            pages: " 10.1126/science.aaw5202. Epub 2019 Jul 18"
COMMENT     Sequence Label: pAAV-hSyn-ChRmine-mScarlet-WPRE
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      653..662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1655..2347
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(1785..1803)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2053..2072
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2399..2987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2452..2472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2870..2881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3019..3495
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3535..3675
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3535..3664
                     /label=AAV2 ITR
     rep_origin      3750..4205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3837..3856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4047..4068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4222..4241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4341..4363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4401..4419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4487..4591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4592..4660,4661..5452)
                     /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(4810..4829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5623..6211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6112..6131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggatcc ccgggtaccg gtgccaccat
      661 ggcacacgca ccaggcaccg accagatgtt ctacgtgggc acaatggacg gctggtatct
      721 ggataccaag ctgaactccg tggccatcgg cgcccactgg tcttgcttta tcgtgctgac
      781 aatcaccaca ttctacctgg gctatgagtc ctggacatct cggggcccta gcaagagaac
      841 ctccttttac gccggctatc aggaggagca gaacctggcc ctgtttgtga atttctttgc
      901 catgctgagc tacttcggca agatcgtggc cgacacactg ggccacaact tcggcgatgt
      961 gggccccttc atcatcggct tcggcaatta caggtatgcc gactacatgc tgacctgccc
     1021 aatgctggtg tacgatctgc tgtatcagct gagggccccc tatcgcgtgt cttgtagcgc
     1081 catcatcttt gccatcctga tgtctggcgt gctggccgag ttctacgcag agggcgaccc
     1141 taggctgagg aatggcgcct acgcctggta tggctttggc tgtttctggt ttatcttcgc
     1201 ctactctatc gtgatgagca tcgtggccaa gcagtatagc cggctggccc agctggccca
     1261 ggatacaggc gccgagcaca gcctgcacgt gctgaagttc gccgtgttta ccttctccat
     1321 gctgtggatt ctgtttcccc tggtgtgggc catctgccct agaggcttcg gctggatcga
     1381 cgataactgg acagaggtgg cccactgcgt gtgcgacatc gtggccaagt cctgttacgg
     1441 ctttgccctg gcccggttca gaaagaccta tgatgaggag ctgtttcggc tgctggagca
     1501 gctgggacac gacgaggatg agttccagaa gctggagctg gatatgaggc tgagcagcaa
     1561 tggcgagcgc ctgcggagac tgtctgcggc cgccaagagc aggatcacca gcgagggcga
     1621 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg cagtgatcaa
     1681 ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat
     1741 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1801 caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc
     1861 cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga
     1921 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca
     1981 ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt
     2041 ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg
     2101 gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga
     2161 cggcgggcgc tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat
     2221 gcccggcgcc tacaacgtgg accgcaagtt ggacatcacc tcccacaacg aggactacac
     2281 cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct
     2341 gtacaagttc tgctacgaga acgaggtgta atgagaattc gatatcaagc ttatcgataa
     2401 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2461 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2521 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2581 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2641 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2701 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2761 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2821 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2881 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2941 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     3001 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     3061 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     3121 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     3181 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     3241 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     3301 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     3361 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     3421 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     3481 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     3541 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3601 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3661 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3721 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3961 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4081 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4201 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6241 cctggccttt tgctggcctt ttgctca
//
