LOCUS       sequence-479180-        6744 bp    DNA     circular SYN 01-AUG-2025
DEFINITION  Cre-dependent expression of LATeNT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  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 6744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 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-DIO-mCherry-LATeNT
FEATURES             Location/Qualifiers
     source          1..6744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2650..2691)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(2692..3399)
                     /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(2769..2792)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2967..2986)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3236..3254
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3396..3405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3413..3446
                     /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    3497..3530
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3600..3620)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4007..4147
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4007..4136
                     /label=AAV2 ITR
     rep_origin      4222..4677
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4309..4328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4519..4540
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4694..4713)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4813..4835
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4873..4891)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4959..5063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5064..5132,5133..5924)
                     /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(5282..5301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6095..6683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6584..6603
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgcctaccg gtactagttc aagcggtacg gttgtacagg ttttcacgga tgttggtcgg
      841 cgggatgatt tttttgcaca gaccgatcag tttggaaacc agaccggaac cgtcgacgtt
      901 cctaaaggcg ttagtgttca ccctcatgtt ctggccctta tactcggact tcagatcctt
      961 ggactctatg ttaaagccct cagtatcgtt atatatagta tcatccagca ggttaggtat
     1021 cttcacagga tcgtggttca tggaaaaata gcttaagcga gtctttatgt taaacttctt
     1081 gcccagctct atctcagtaa aaatctgaaa ggctgtcttt ttgaccaggc agaccgcctc
     1141 acgctcagcg gcgccatgaa ggcgttcggt gccatccagc tgcacgccaa tgaagtactg
     1201 gacgtcgccc ttataatcgc gcatcggctg caggtggaac acgttccaga acttcttgcc
     1261 gctcttggta taattgatca gctgaacggt cacttcagtc tggttgtcaa tagcatctct
     1321 gattttccga acagtggcac ggtctgtctc gggtccctgc agaaaacggc aattgcgccc
     1381 caggatctct tcccgagaat attctgtcag ctgcaggaag gaattgctca caaaaatgat
     1441 tgggttgtca ggcagtctcg gatcagtaat cacgaagctt ttctcgatcc gttccagtgt
     1501 ggttgcccga aactccccgc taccggaccc cccgctgccg ctcccgccat acattatgga
     1561 gttatacagt atctgaaact tatcctcgtt cactatatac tggccgttgg aatccttatc
     1621 aaactgatac ttctgctgat agatctgttt gtaggagtcg atgtcgatgt tcgggtcgtt
     1681 gcaggaggta acctgggaca gtttgttagc gatagctttg tagtcgttca gggttttttc
     1741 gtacaggtcg tttttgatat cgatggagat caggttagcg tcctgaccac cgaaggtgaa
     1801 gagctcttca gcggagatcg ggtaggtgtg ctgcatgtag atttcctgtt tggacgggat
     1861 gatttcgtgg gaggaaacct gcatgccgta cagaccgtgc agaacgtgga tcagttcgtg
     1921 catcagcagc agagccgggt cctggaagta tttggattta ccgatggtca gggaggtgat
     1981 gttttcgata acgttgtcga aggtcggtac gtattccggg cagaaagcca tctgcatgat
     2041 ggaaccgaaa ccgtcacggc acgggaagta gtttttgttg tcaacacgca gaacgatacc
     2101 acgaacttcg tttttgttca gaaccggacc cggaccgaag atgatcaggt tggtcagcat
     2161 agcggatttg gtggtagcac cggacggatc ctgttccagc aggttgaagg aaacggagtt
     2221 ggagttggtg tcgaatttgt ccagcaggga gtaggagtta cccaggtacg ggatagcgtt
     2281 gatgattttg tccagcagag cttcaccagc aacgttgttt ttgatacggt tgaacagttt
     2341 aaccatggtc tgcaggaaac ggtctttgtc ggagtcggta cgcaggtagt tcgggtcgta
     2401 gtattcggaa gcaccttcga tcagggagga cggcgggttg aagtcttccg gtttggtacc
     2461 aaattcgtaa cgttccggaa cgatccagat acggtcggtg attttgaagg ctctgtagta
     2521 gatgtctaga cctttgcagt acggcggttc catcatgatg atggtgtcgt tgttaaccgg
     2581 gtcggagtaa cggaagttgt tgatggtgat cggaccggtg cccgagcccg aggtcgagcc
     2641 cgagcccttg gtgctgtcca ggcccagcag ggggttgggg atgggcttgc ccttgtacag
     2701 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgc tcgtactgtt ccacgatggt
     2761 gtagtcctcg ttgtgggagg tgatgtccag cttgatgttg acgttgtagg cgccgggcag
     2821 ctgcacgggc ttcttggcct tgtaggtggt cttgacctcg gcgtcgtagt ggccgccgtc
     2881 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     2941 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     3001 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     3061 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     3121 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     3181 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cgcccttggt
     3241 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     3301 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     3361 gatgatggcc atgttgtcct cctcgccctt gctcaccatg gtggcggcta gcataacttc
     3421 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     3481 tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat gaattccgct
     3541 cgagataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3601 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3661 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     3721 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3781 aattccgtgg tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag
     3841 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3901 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3961 ggttttttaa agcggccgca ggaacacgtg cggaccgagc ggccgcagga acccctagtg
     4021 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4081 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4141 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4201 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4261 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4321 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4381 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4441 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4501 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4561 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4621 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4681 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4741 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4801 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4861 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4921 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4981 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5041 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5101 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5161 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5221 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5281 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5341 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5401 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5461 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5521 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5581 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5641 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5701 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5761 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5821 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5881 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5941 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6001 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6061 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6121 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6181 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6241 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6301 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6361 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6421 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6481 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6541 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6601 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6661 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6721 tttgctggcc ttttgctcac atgt
//
