LOCUS       sequence-235837-        7630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TVA, V5, and RG in a cre dependent manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  AUTHORS   Ahrlund-Richter S, Xuan Y, van Lunteren JA, Kim H, Ortiz C, Pollak 
            Dorocic I, Meletis K, Carlen M
  TITLE     A whole-brain atlas of monosynaptic input targeting four different 
            cell types in the medial prefrontal cortex of the mouse.
  JOURNAL   Nat Neurosci. 2019 Apr;22(4):657-668. doi: 
            10.1038/s41593-019-0354-y. Epub 2019 Mar 18.
  PUBMED    30886408
REFERENCE   2  (bases 1 to 7630)
  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"; doi: 
            "10.1038/s41593-019-0354-y"; journalName: "Nat Neurosci"; date: 
            "2019-04"; volume: "22"; issue: "4"; pages: "657-668"
COMMENT     Sequence Label: pAAV-DIO-TVA-V5-RG
FEATURES             Location/Qualifiers
     source          1..7630
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3138..3191)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(3201..3242)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      3599..3608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3615..3648
                     /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    3699..3732
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3757..4345
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3810..3830)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4228..4239)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4377..4853
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4576..4595)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4893..5033
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4893..5022
                     /label=AAV2 ITR
     rep_origin      5108..5563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5195..5214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5405..5426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5580..5599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5699..5721
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5759..5777)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5845..5949
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5950..6018,6019..6810)
                     /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(6168..6187)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6981..7569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7470..7489
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttaca gtctggtctc acccccactc ttgtgtgatt cccatgaaga tatgatcttc
     1621 ccgctttggg gagtgactga cacctccctc cctgtccctc tgagattgtg ttgcgtaggt
     1681 tctgatcgat tgactcttct acaacatgtc atcaggaaaa ttatcaacat caaggcagtc
     1741 agggcccctg cactcagtaa tacatacttc ccccagttcg ggagacccaa gtcaactcct
     1801 gagacctgat tgtgcacatc gggaaggtga acttcaacaa aatcctcagc ctcgtcaccg
     1861 tccttgaaaa cggtagacgg gtctgccagg gggtgcacaa gggggataac cgaggattcc
     1921 aacaactcca tatgttgctg gaggagggat gattgcatct ctgggattaa gacattgccg
     1981 tcaggtccta atattatacc attgaaaaac accccgttca catgaggatg acacctcccc
     2041 ccaactctta aacacccttt tgaagggagg atctcattcc aagttctgac tgacttgtag
     2101 tgagcatcgg cttccatcaa ggtcttgttg aatatggtat atgcttttcc aaacccaggg
     2161 acaagttttc ttaaatgact gagacgtctg aaactcactg acttggttgt catgatggac
     2221 tctagtgcat ccagacactc ctctctcttc ctgaccaact cctctacaac aaggtgctca
     2281 atttcgtctg agcgaaagtc gtgcaggttc accaacttat cgggagggca ccatttggtt
     2341 tcatttgatg tttgcatcga gacccatgtt ccatccataa gtctaagtcc tagaactcca
     2401 cataacttga gtttgcatgc tccttttaaa gacttatata ggcctctttc atctacaaag
     2461 ccgcaagtct cactcccttt ggatgctctc ttccctctac tattggtaaa aatgtcacaa
     2521 gacatcccta gtctcggatt ctcgggcatc caaatggtgt aatcgtggtt agtggagcag
     2581 taggtagaag acaccgctac tcctgagcac ttcccgctag ggaagaccct cgagtgaagg
     2641 gatctgtcat atgggtccaa atctgccaca cttggagata tgataacgag agactccttg
     2701 gtggttttta cagttcgaag ccagcggtag tcagggtacg gattgtgtag agactcttca
     2761 tatctggggt caccggccat cttccagttg tacgcggctc tacatgcatc tggtgttggg
     2821 cggaaatgct ttcttttgaa cgtggttgtg acataaccaa cgaagttagt gtaggtttca
     2881 gcctccgtca caacgcctgt gcaagtgaac ccgttcactt ttatggctaa gatgtatcca
     2941 actttaagtt ccatgtagga gaaccctgac aggttggtgc atccttcgtc ctccactacc
     3001 aaattgtttg ggcagctgag gtgatgtatg tcaatcggac tccagggacc aagcttgtct
     3061 ggtatcgtgt aaatagggaa tttcccaaaa cacaatggaa aaaccagaag gggtacaaac
     3121 aggagagcct gaggaactgg gccaggattc tcctcgacgt caccgcatgt tagcagactt
     3181 cctctgccct ctccactgcc cgtagaatcg agaccgagga gagggttagg gataggctta
     3241 ccgtcccatc tcaccagctc acagcaaaag atccctgcag tgatcagcat ccacatgcgg
     3301 ccgtgattcc tggctggcag agcacgtcca ggagcaggga cagtgggagc ctctgtgccg
     3361 ttgtccgtgg gcaccgcggg ggtcgcgctg gtcccgcagc cccactcgtc ccgcccgtcg
     3421 tcgcagtcgg ggtgtccgtc gcacagccag tcctgcgggt aacactcccc gtgggcaccg
     3481 ggcggctccg agcagcggaa ctgaccgggg gggcaacggg acaaagaacc gttaccggac
     3541 ccgttaccgg tcacgttacc gggcagcagc agcagcagca gcgcgggcag cagccgcgcc
     3601 atggtggcgc tagcataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     3661 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aatgtatgct
     3721 atacgaagtt atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3781 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3841 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3901 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3961 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4021 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4081 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4141 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     4201 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4261 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4321 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     4381 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     4441 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     4501 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     4561 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     4621 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     4681 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     4741 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     4801 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     4861 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     4921 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4981 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     5041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     5101 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5161 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     5221 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5281 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     5341 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5401 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     5461 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     5521 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     5581 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5641 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5701 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5761 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5821 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5881 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5941 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6001 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6061 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6121 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6181 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6241 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6301 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6361 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6421 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6481 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6541 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6601 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6661 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6721 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6781 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6841 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6901 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6961 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7021 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7081 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7141 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7201 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7261 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7321 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7381 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7441 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7501 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7561 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7621 gctcacatgt
//
