LOCUS       sequence-479179-        6706 bp    DNA     circular SYN 08-AUG-2025
DEFINITION  Expresses mCherry-TeNT in mammalian cells; for AAV transduction of 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   Roh H, Kim D, Kim B, Jeon Y, Kim Y, Jacko M, Xu F, Lin C, Won Um J, 
            Ting AY
  TITLE     Light-activated tetanus neurotoxin for conditional proteolysis and 
            inducible synaptic inhibition in vivo
  JOURNAL   bioRxiv 2025.01.27.635161; doi: 10.1101/2025.01.27.635161
REFERENCE   2  (bases 1 to 6706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.01.27.635161; doi: 10.1101/2025.01.27.635161"
COMMENT     Sequence Label: pAAV-hSyn-mCherry-TeNT
FEATURES             Location/Qualifiers
     source          1..6706
                     /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             659..1366
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(804..822)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1072..1091
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1266..1289
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1367..1408
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2838..3426
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2891..2911)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3309..3320)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3458..3934
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3657..3676)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3974..4114
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3974..4103
                     /label=AAV2 ITR
     rep_origin      4189..4644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4276..4295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4486..4507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4661..4680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4780..4802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4840..4858)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4926..5030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5031..5099,5100..5891)
                     /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(5249..5268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6062..6650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6551..6570
                     /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 agaaggtacc ggatctgcta gcgccaccat
      661 ggtgagcaag ggcgaggagg acaacatggc catcatcaag gagttcatgc gcttcaaggt
      721 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
      781 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
      841 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
      901 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
      961 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     1021 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     1081 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     1141 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgccga
     1201 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     1261 catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     1321 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggca agcccatccc
     1381 caaccccctg ctgggcctgg acagcaccaa gggctcgggc tcgacctcgg gctcgggcac
     1441 cggtccgatc accatcaaca acttccgtta ctccgacccg gttaacaacg acaccatcat
     1501 catgatggaa ccgccgtact gcaaaggtct agacatctac tacaaagcct tcaaaatcac
     1561 cgaccgtatc tggatcgttc cggaacgtta cgaatttggt accaaaccgg aagacttcaa
     1621 cccgccgtcc tccctgatcg aaggtgcttc cgaatactac gacccgaact acctgcgtac
     1681 cgactccgac aaagaccgtt tcctgcagac catggttaaa ctgttcaacc gtatcaaaaa
     1741 caacgttgct ggtgaagctc tgctggacaa aatcatcaac gctatcccgt acctgggtaa
     1801 ctcctactcc ctgctggaca aattcgacac caactccaac tccgtttcct tcaacctgct
     1861 ggaacaggat ccgtccggtg ctaccaccaa atccgctatg ctgaccaacc tgatcatctt
     1921 cggtccgggt ccggttctga acaaaaacga agttcgtggt atcgttctgc gtgttgacaa
     1981 caaaaactac ttcccgtgcc gtgacggttt cggttccatc atgcagatgg ctttctgccc
     2041 ggaatacgta ccgaccttcg acaacgttat cgaaaacatc acctccctga ccatcggtaa
     2101 atccaaatac ttccaggacc cggctctgct gctgatgcac gaactgatcc acgttctgca
     2161 cggtctgtac ggcatgcagg tttcctccca cgaaatcatc ccgtccaaac aggaaatcta
     2221 catgcagcac acctacccga tctccgctga agagctcttc accttcggtg gtcaggacgc
     2281 taacctgatc tccatcgata tcaaaaacga cctgtacgaa aaaaccctga acgactacaa
     2341 agctatcgct aacaaactgt cccaggttac ctcctgcaac gacccgaaca tcgacatcga
     2401 ctcctacaaa cagatctatc agcagaagta tcagtttgat aaggattcca acggccagta
     2461 tatagtgaac gaggataagt ttcagatact gtataactcc ataatgtatg gctttactga
     2521 gatagagctg ggcaagaagt ttaacataaa gactcgctta agctattttt ccatgaacca
     2581 cgatcctgtg aagataccta acctgctgga tgatactata tataacgata ctgagggctt
     2641 taacatagag tccaaggatc tgaagtccga gtataagggc cagaacatga gggtgaacac
     2701 taacgccttt aggaacgtcg acggttccgg tctggtttcc aaactgatcg gtctgtgcaa
     2761 aaaaatcatc ccgccgacca acatccgtga aaacctgtac aaccgtaccg cttgaattcg
     2821 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2881 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2941 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3001 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3061 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3121 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3181 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3241 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3301 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3361 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3421 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3481 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3541 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3601 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3661 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3721 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3781 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3841 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3901 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3961 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4021 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4081 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4141 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4201 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4261 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4321 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4381 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4441 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4501 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4561 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4621 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4681 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4741 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4801 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4861 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5041 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5101 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5161 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5221 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5281 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5341 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5401 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5461 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5521 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5581 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5641 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5701 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5761 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5821 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5881 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6181 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctca
//
