LOCUS       sequence-361663-        7307 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cre-dependent AAV virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7307)
  AUTHORS   Yao S, Wang Q, Hirokawa KE, Ouellette B, Ahmed R, Bomben J, Brouner 
            K, Casal L, Caldejon S, Cho A, Dotson NI, Daigle TL, Egdorf T, 
            Enstrom R, Gary A, Gelfand E, Gorham M, Griffin F, Gu H, Hancock N, 
            Howard R, Kuan L, Lambert S, Lee EK, Luviano J, Mace K, Maxwell M, 
            Mortrud MT, Naeemi M, Nayan C, Ngo NK, Nguyen T, North K, Ransford 
            S, Ruiz A, Seid S, Swapp J, Taormina MJ, Wakeman W, Zhou T, Nicovich
            PR, Williford A, Potekhina L, McGraw M, Ng L, Groblewski PA, Tasic 
            B, Mihalas S, Harris JA, Cetin A, Zeng H
  TITLE     A whole-brain monosynaptic input connectome to neuron classes in 
            mouse visual cortex.
  JOURNAL   Nat Neurosci. 2023 Feb;26(2):350-364. doi: 
            10.1038/s41593-022-01219-x. Epub 2022 Dec 22.
  PUBMED    36550293
REFERENCE   2  (bases 1 to 7307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41593-022-01219-x"; journalName: "Nat Neurosci"; date: 
            "2023-02"; volume: "26"; issue: "2"; pages: "350-364"
COMMENT     Sequence Label: AiP5005 pAAV-Syn-DIO-TVA66T-dTom-SAD-B19G
FEATURES             Location/Qualifiers
     source          1..7307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(32..620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(112..131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(791..1582,1583..1651))
                     /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     1414..1433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1652..1756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1824..1842
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1880..1902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2002..2021
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2038..2493)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2175..2196)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2387..2406
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2568..2708
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2579..2708
                     /label=AAV2 ITR
     polyA_signal    2710..2934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    2942..3530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3048..3059
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3457..3477
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3532..3565
                     /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    3616..3649
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4132..4188
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4189..4887
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(4319..4337)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4587..4606
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4781..4804
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4897..4953
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(6534..6567)
                     /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(6618..6651)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      6657..6666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6673..7120)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   7148..7277
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
       61 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      121 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      181 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      241 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      301 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      361 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      421 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      481 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      541 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      601 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      661 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      721 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
      781 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      841 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      901 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      961 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1021 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1081 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1141 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1201 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1261 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1321 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1381 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1441 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1501 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1561 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1621 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1681 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1741 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     1801 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     1861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1981 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa
     2041 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2101 ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
     2161 agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2221 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
     2281 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2341 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2401 aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2461 ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg
     2521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct
     2581 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2641 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2701 gggttcctgc catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct
     2761 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
     2821 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
     2881 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acaggatcca
     2941 ggcggggagg cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga
     3001 agaggccgca gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa
     3061 gggacgtagc agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccat
     3121 ggaaaggacg tcagcttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc
     3181 cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg
     3241 aaagcgaaag tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt
     3301 gcgtcagcaa acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca
     3361 taaagagaca gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca
     3421 atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag
     3481 ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt cataacttcg
     3541 tataaagtat cctatacgaa gttattcttt gcaccattct aaagaataac agtgataatt
     3601 tctgggttaa ggcaaataac ttcgtataat gtatgctata cgaagttatt aggtgcgcgg
     3661 ctgctgcccg cgctgctgct gctgctgctg cccggtaacg tgaccggtaa cgggtccggt
     3721 aacggttctt tgtcccgttg cccccccggt cagttccgct gctcggagcc gcccggtgcc
     3781 cacggggagt gttacccgca ggactggctg tgcgacggac accccgactg cgacgacggg
     3841 cgggacacct ggggctgcgg gaccagcgcg atccccgcgg tgcccacgga caacggcaca
     3901 gaggctccca ctgcccctgc tcctggacgt gctctgccag ccaggaatca cggccgcatg
     3961 tggatgctga tcactgcagt gctcctgtgc tgcctggtag ctgtgggtgg tatcgctgca
     4021 tgggggaagt ccaaagcaaa aagcaggtct gacatcttca gtcttgaaag cgcatccaag
     4081 gagctgctgg tgcctgacaa gagccaggca gacttgttct ccggatccgg agctactaac
     4141 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacccgt gagcaagggc
     4201 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     4261 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     4321 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     4381 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     4441 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     4501 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     4561 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     4621 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     4681 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     4741 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     4801 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     4861 ctgtacggca tggacgagct gtacaaggga tccggagcta ctaacttcag cctgctgaag
     4921 caggctggag acgtggagga gaaccctgga cccatggttc ctcaggctct cctgtttgta
     4981 ccccttctgg tttttccatt gtgttttggg aaattcccta tttacacgat accagacaag
     5041 cttggtccct ggagtccgat tgacatacat cacctcagct gcccaaacaa tttggtagtg
     5101 gaggacgaag gatgcaccaa cctgtcaggg ttctcctaca tggaacttaa agttggatac
     5161 atcttagcca taaaagtgaa cgggttcact tgcacaggcg ttgtgacgga ggctgaaacc
     5221 tacactaact tcgttggtta tgtcacaacc acgttcaaaa gaaagcattt ccgcccaaca
     5281 ccagatgcat gtagagccgc gtacaactgg aagatggccg gtgaccccag atatgaagag
     5341 tctctacaca atccgtaccc tgactaccgc tggcttcgaa ctgtaaaaac caccaaggag
     5401 tctctcgtta tcatatctcc aagtgtggca gatttggacc catatgacag atcccttcac
     5461 tcgagggtct tccctagcgg gaagtgctca ggagtagcgg tgtcttctac ctactgctcc
     5521 actaaccacg attacaccat ttggatgccc gagaatccga gactagggat gtcttgtgac
     5581 atttttacca atagtagagg gaagagagca tccaaaggga gtgagacttg cggctttgta
     5641 gatgaaagag gcctatataa gtctttaaaa ggagcatgca aactcaagtt atgtggagtt
     5701 ctaggactta gacttatgga tggaacatgg gtctcgatgc aaacatcaaa tgaaaccaaa
     5761 tggtgccctc ccgataagtt ggtgaacctg cacgactttc gctcagacga aattgagcac
     5821 cttgttgtag aggagttggt caggaagaga gaggagtgtc tggatgcact agagtccatc
     5881 atgacaacca agtcagtgag tttcagacgt ctcagtcatt taagaaaact tgtccctggg
     5941 tttggaaaag catataccat attcaacaag accttgatgg aagccgatgc tcactacaag
     6001 tcagtcagaa cttggaatga gatcctccct tcaaaagggt gtttaagagt tggggggagg
     6061 tgtcatcctc atgtgaacgg ggtgtttttc aatggtataa tattaggacc tgacggcaat
     6121 gtcttaatcc cagagatgca atcatccctc ctccagcaac atatggagtt gttggaatcc
     6181 tcggttatcc cccttgtgca ccccctggca gacccgtcta ccgttttcaa ggacggtgac
     6241 gaggctgagg attttgttga agttcacctt cccgatgtgc acaatcaggt ctcaggagtt
     6301 gacttgggtc tcccgaactg ggggaagtat gtattactga gtgcaggggc cctgactgcc
     6361 ttgatgttga taattttcct gatgacatgt tgtagaagag tcaatcgatc agaacctacg
     6421 caacacaatc tcagagggac agggagggag gtgtcagtca ctccccaaag cgggaagatc
     6481 atatcttcat gggaatcaca caagagtggg ggtgagacca gactgtaggt accataactt
     6541 cgtataggat actttatacg aagttattaa gaggtttcat attgctaata gcagctacaa
     6601 tccagctacc attctgcata acttcgtata gcatacatta tacgaagtta tattaaccat
     6661 ggtggcggct gactgcgctc tcaggcacga cacgactcct ccgctgccca ccgcagactg
     6721 aggcagcgct gagtcgccgg cgccgcagcg cagatggtcg cgcccgtgcc cccctatctc
     6781 gcgcctcgcg tggtgcggtc cggctgggcc ggcggcggcg cggacgcgac caaggtggcc
     6841 gggaagggga gtttgcgggg gaccggcgag tgacgtcagc gcgccttcag tgctgaggcg
     6901 gcggtggcgc gcgccgccag gcgggggcga aggcactgtc cgcggtgctg aagctggcag
     6961 tgcgcacgcg cctcgccgca tcctgtttcc cctccccctc tctgataggg gatgcgcaat
     7021 ttggggaatg ggggttgggt gcttgtccag tgggtcgggg tcggtcgtca ggtaggcacc
     7081 cccaccccgc ctcatcctgg tcctaaaacc cacttgcact catacgcagg gccctctgca
     7141 gtctagcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     7201 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     7261 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggcca
//
