LOCUS       sequence-470281-        6312 bp    DNA     circular SYN 28-APR-2025
DEFINITION  AiE0454m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, Weed N, Wirthlin M, Wood 
            T, Wynalda B, Yao Z, Zhou T, Ariza J, Dee N, Reding M, Ronellenfitch
            K, Mufti S, Sunkin SM, Smith KA, Esposito L, Waters J, Thyagarajan 
            B, Yao S, Lein ES, Zeng H, Levi BP, Ngai J, Ting J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 6312)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1188 - pAAV-AiE0454m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1110..1119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1116..1121,1122..1142,1143..2414)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    2439..3027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2492..2512)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2910..2921)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3059..3535
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3258..3277)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3575..3715
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3575..3704
                     /label=AAV2 ITR
     rep_origin      3790..4245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4087..4108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4262..4281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4381..4403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4441..4459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4527..4631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4632..4700,4701..5492)
                     /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(4850..4869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5663..6251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6152..6171
                     /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 gcgtagaacc aagtgcccga ccccagagtc tttctttgtc
      181 tgacctgagg gctttctgtt aaaatcctta tacagcagat aagtaagtca gcagtcttag
      241 aggtgggaag aggtctgtct gccagtcatc tggctccctg caaacacagg catccccacc
      301 cctgaacaat tttcttcata ggaaagttgt gagggaagtc acaggtgcag acaccgcccc
      361 ggggggggag ggggggctca gagttgttcc ttcattcagt caccaattca gtcttccaga
      421 atcaaggccc tattgacaga gggctaagcc accggggcgc tgccttcaag gtaagaggca
      481 tagatcagtc tcctgaatgt agagggatga attccctcct tcacataagt gaaagcgatc
      541 atagggaacg gaggcatcat tagggcgccc tccttctgac agggatgcag acaggcaggt
      601 gagggtggag gggtgtggcc cagggcacat tcagagccaa actctgttga gaacaaggca
      661 ggtgtttctg aaaacctctc tgattgcacc cgtaccctcc catcaggccc gaatcgcaca
      721 cctctctgcc tctcctcaga cgccggccca catggctttt gaaaacctgg ggagattata
      781 ccttccagct tgctttgctg agtctgagga acctggctgc cttttctctt ctgttaactg
      841 caatcaaact ggttttgagc accagcccca agatgtcttt gccagtccac atccatcttc
      901 catgatgagc taaggttccc tatttgtacc cttactttct gggctgagtc atgacatctg
      961 ggaatggtgc tttctctgag aacttgcggc ggcggcgggg ggggggggga gtggctgagg
     1021 gggggttacg aagacaatgc ttaaggagct cgggctgggc ataaaagtca gggcagagcc
     1081 atctattgct tacatttgct tctggcgtgg ccaccatggc tcctaagaag aagaggaagg
     1141 tgatgagcca gttcgacatc ctgtgcaaga ccccccccaa ggtgctggtg cggcagttcg
     1201 tggagagatt cgagaggccc agcggcgaga agatcgccag ctgtgccgcc gagctgacct
     1261 acctgtgctg gatgatcacc cacaacggca ccgccatcaa gagggccacc ttcatgagct
     1321 acaacaccat catcagcaac agcctgagct tcgacatcgt gaacaagagc ctgcagttca
     1381 agtacaagac ccagaaggcc accatcctgg aggccagcct gaagaagctg atccccgcct
     1441 gggagttcac catcatccct tacaacggcc agaagcacca gagcgacatc accgacatcg
     1501 tgtccagcct gcagctgcag ttcgagagca gcgaggaggc cgacaagggc aacagccaca
     1561 gcaagaagat gctgaaggcc ctgctgtccg agggcgagag catctgggag atcaccgaga
     1621 agatcctgaa cagcttcgag tacaccagca ggttcaccaa gaccaagacc ctgtaccagt
     1681 tcctgttcct ggccacattc atcaactgcg gcaggttcag cgacatcaag aacgtggacc
     1741 ccaagagctt caagctggtg cagaacaagt acctgggcgt gatcattcag tgcctggtga
     1801 ccgagaccaa gacaagcgtg tccaggcaca tctacttttt cagcgccaga ggcaggatcg
     1861 accccctggt gtacctggac gagttcctga ggaacagcga gcccgtgctg aagagagtga
     1921 acaggaccgg caacagcagc agcaacaagc aggagtacca gctgctgaag gacaacctgg
     1981 tgcgcagcta caacaaggcc ctgaagaaga acgcccccta ccccatcttc gctatcaaga
     2041 acggccctaa gagccacatc ggcaggcacc tgatgaccag ctttctgagc atgaagggcc
     2101 tgaccgagct gacaaacgtg gtgggcaact ggagcgacaa gagggcctcc gccgtggcca
     2161 ggaccaccta cacccaccag atcaccgcca tccccgacca ctacttcgcc ctggtgtcca
     2221 ggtactacgc ctacgacccc atcagcaagg agatgatcgc cctgaaggac gagaccaacc
     2281 ccatcgagga gtggcagcac atcgagcagc tgaagggcag cgccgagggc agcatcagat
     2341 accccgcctg gaacggcatc atcagccagg aggtgctgga ctacctgagc agctacatca
     2401 acaggcggat ctgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3001 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     3061 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3121 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3181 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3241 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3301 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     3361 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     3421 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3481 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3541 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3601 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3661 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3721 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3781 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3841 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4141 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4201 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6301 ttgctcacat gt
//
