LOCUS       sequence-478938-        7442 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses NanoLuc-LATeNT* in neurons; for AAV transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7442)
  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 7442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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-CAG-NanoLuc-LATeNT(slow)
FEATURES             Location/Qualifiers
     source          1..7442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        216..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..797
                     /label=chicken beta-actin promoter
     intron          798..1814
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1738..1760
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1827..1836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1836..1877
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1914..2423
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4330..4350)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4705..4845
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4705..4834
                     /label=AAV2 ITR
     rep_origin      4920..5375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5007..5026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5217..5238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5392..5411)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5511..5533
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5571..5589)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5657..5761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5762..5830,5831..6622)
                     /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(5980..5999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6793..7381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7282..7301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctgcag gaatctagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      541 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      601 taattatttt gtgcagcgat gggggcgggg gggggggggg gcgcgcgcca ggcggggcgg
      661 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
      721 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      781 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      841 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      901 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
      961 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
     1021 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
     1081 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1141 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1201 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1261 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1321 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1381 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     1441 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1501 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1561 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1621 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1681 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     1741 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     1801 tctttttcct acagcaattg gcggccgcca ccatgggcaa gcccatcccc aaccccctgc
     1861 tgggcctgga cagcaccaag ggctcgggct cgacctcggg ctcgggcacc ggtgtcttca
     1921 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     1981 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     2041 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     2101 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc
     2161 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg
     2221 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     2281 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     2341 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc
     2401 ggctgtgcga acgcattctg gcggggagcg gcagcggggg gtccggtagc gggccgatca
     2461 ccatcaacaa cttccgttac tccgacccgg ttaacaacga caccatcatc atgatggaac
     2521 cgccgtactg caaaggtcta gacatctact acagagcctt caaaatcacc gaccgtatct
     2581 ggatcgttcc ggaacgttac gaatttggta ccaaaccgga agacttcaac ccgccgtcct
     2641 ccctgatcga aggtgcttcc gaatactacg acccgaacta cctgcgtacc gactccgaca
     2701 aagaccgttt cctgcagacc atggttaaac tgttcaaccg tatcaaaaac aacgttgctg
     2761 gtgaagctct gctggacaaa atcatcaacg ctatcccgta cctgggtaac tcctactccc
     2821 tgctggacaa attcgacacc aactccaact ccgtttcctt caacctgctg gaacaggatc
     2881 cgtccggtgc taccaccaaa tccgctatgc tgaccaacct gatcatcttc ggtccgggtc
     2941 cggttctgaa caaaaacgaa gttcgtggta tcgttctgcg tgttgacaac aaaaactact
     3001 tcccgtgccg tgacggtttc ggttccatca tgcagatggc tttctgcccg gaatacgtac
     3061 cgaccttcga caacgttatc gaaaacatca cctccctgac catcggtaaa tccaaatact
     3121 tccaggaccc ggctctgctg ctgatgcacg aactgatcca cgttctgcac ggtctgtacg
     3181 gcatgcaggt ttcctcccac gaaatcatcc cgtccaaaca ggaaatctac atgcagcaca
     3241 cctacccgat ctccgctgaa gagctcttca ccttcggtgg tcaggacgct aacctgatct
     3301 ccatcgatat caaaaacgac ctgtacgaaa aaaccctgaa cgactacaaa gctatcgcta
     3361 acaaactgtc ccaggttacc tcctgcaacg acccgaacat cgacatcgac tcctacaaac
     3421 agatctatca gcagaagtat cagtttgata aggattccaa cggccagtat atagtgaacg
     3481 aggataagtt tcagatactg tataactcca taatgtatgg cgggagcggc agcggggggt
     3541 ccggtagcgg ggagtttcgg gcaaccacac tggaacggat cgagaaaagc ttcctgatta
     3601 ctgatccgag actgcctgac aacccaatca tttttgtgag caattccttc ctgcagctga
     3661 cagaatattc tcgggaagag atcctggggc gcaattgccg ttttctgcag ggacccgaga
     3721 cagaccgtgc cactgttcgg aaaatcagag atgctattga caaccagact gaagtgaccg
     3781 ttcagctgat caattatacc aagagcggca agaagttctg gaacgtgttc cacctgcagc
     3841 cgatgcgcga ttataagggc gacgtccagt acttcattgg cgtgcagctg gatggcaccg
     3901 aacgccttca tggcgccgct gagcgtgagg cggtctgcct ggtcaaaaag acagcctttc
     3961 agatttttac tgagatagag ctgggcaaga agtttaacat aaagactcgc ttaagctatt
     4021 tttccatgaa ccacgatcct gtgaagatac ctaacctgct ggatgatact atatataacg
     4081 atactgaggg ctttaacata gagtccaagg atctgaagtc cgagtataag ggccagaaca
     4141 tgagggtgaa cactaacgcc tttaggaacg tcgacggttc cggtctggtt tccaaactga
     4201 tcggtctgtg caaaaaaatc atcccgccga ccaacatccg tgaaaacctg tacaaccgta
     4261 ccgcttaaac tagtataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4321 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4381 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     4441 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4501 gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg ctttatttgt
     4561 aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4621 gctgcaataa acaagttaac aacaacattg cattcatttt atgtttcagg ttcaggggga
     4681 gatgtgggaa ctagttgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4741 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4801 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4861 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5221 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5281 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5341 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5401 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5461 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5521 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5581 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5641 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5701 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5761 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5821 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5881 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5941 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6001 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6061 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6121 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6181 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6241 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6301 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6361 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6421 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6481 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6541 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6601 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6661 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6721 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7441 gt
//
