LOCUS       sequence-466213-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2055m 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 6027)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, 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 JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 6027)
  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"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP1216 - pAAV-AiE2055m-minBG-FlpO-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   444..573
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(635..1223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(715..734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1394..2185,2186..2254))
                     /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     2017..2036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2255..2359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2427..2445
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2483..2505)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2605..2624
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2641..3096)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2799)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2990..3009
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3171..3311
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3182..3311
                     /label=AAV2 ITR
     polyA_signal    3351..3827
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3609..3628
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3859..4447
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3965..3976
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4374..4394
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(4472..5743,5744..5764,5765..5770))
                     /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"
     regulatory      5767..5776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgagcccatg ctgggaattt gaccttcact gtgtgggcag ttgtgtgtgt gtggggggcg
       61 ggggagtcac tgtggaattt gtggtttaca ttttgaaagt gtctctctag tagcaaggtg
      121 agaaactgac aaaagtcact ctaaagagat ggaaggaaag gcaggaaagc acttaacagc
      181 tccagctaac acttaccaac tccagtagtt gagccaaagg ccatgtgaat gggctgagtc
      241 atctgtcctg tcttccccct acccatcatg tcctttctag actcagtacc atttcagatg
      301 atatttctgt ctctggaatt cctttccttc tttcgtcact taaacccttc tacccttctg
      361 agttccattc tgtcgtcccg tctctaagaa aacctgtgct ataagtcgtc ttacacagga
      421 ggacttagca cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
      481 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
      541 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
      601 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      661 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      721 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      781 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      841 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      901 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      961 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1021 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1081 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1141 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1201 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1261 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1321 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1381 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1441 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1501 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1561 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1621 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1681 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1741 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1801 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1861 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1921 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1981 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2041 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2101 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2161 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2221 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2281 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2341 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2401 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2461 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2521 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2581 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2641 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2701 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     2761 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2821 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     2881 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2941 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3001 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3061 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
     3121 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
     3181 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     3241 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     3301 taggggttcc tgcggccgct cggtccgcac gtggttacct acaaaatcag aaggacaggg
     3361 aagggagcag tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc
     3421 acctgagatt aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa
     3481 aaaaacaaaa attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc
     3541 tgaggcagga gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc
     3601 actgcactcc agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact
     3661 tgccccttgc tccataccac ccccctccac cccataatat tatagaagga cacctagtca
     3721 gacaaaatga tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact
     3781 tccagggcca ggagaggcac tggggagggg tcacagggat gccacccgta gatctctcga
     3841 gcagcgctcg gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc
     3901 gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg
     3961 ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa
     4021 cataggcgag cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag
     4081 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
     4141 tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc
     4201 cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa
     4261 cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga
     4321 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
     4381 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     4441 gttgattatc gataagcttg atatcgaatt ctcagatccg cctgttgatg tagctgctca
     4501 ggtagtccag cacctcctgg ctgatgatgc cgttccaggc ggggtatctg atgctgccct
     4561 cggcgctgcc cttcagctgc tcgatgtgct gccactcctc gatggggttg gtctcgtcct
     4621 tcagggcgat catctccttg ctgatggggt cgtaggcgta gtacctggac accagggcga
     4681 agtagtggtc ggggatggcg gtgatctggt gggtgtaggt ggtcctggcc acggcggagg
     4741 ccctcttgtc gctccagttg cccaccacgt ttgtcagctc ggtcaggccc ttcatgctca
     4801 gaaagctggt catcaggtgc ctgccgatgt ggctcttagg gccgttcttg atagcgaaga
     4861 tggggtaggg ggcgttcttc ttcagggcct tgttgtagct gcgcaccagg ttgtccttca
     4921 gcagctggta ctcctgcttg ttgctgctgc tgttgccggt cctgttcact ctcttcagca
     4981 cgggctcgct gttcctcagg aactcgtcca ggtacaccag ggggtcgatc ctgcctctgg
     5041 cgctgaaaaa gtagatgtgc ctggacacgc ttgtcttggt ctcggtcacc aggcactgaa
     5101 tgatcacgcc caggtacttg ttctgcacca gcttgaagct cttggggtcc acgttcttga
     5161 tgtcgctgaa cctgccgcag ttgatgaatg tggccaggaa caggaactgg tacagggtct
     5221 tggtcttggt gaacctgctg gtgtactcga agctgttcag gatcttctcg gtgatctccc
     5281 agatgctctc gccctcggac agcagggcct tcagcatctt cttgctgtgg ctgttgccct
     5341 tgtcggcctc ctcgctgctc tcgaactgca gctgcaggct ggacacgatg tcggtgatgt
     5401 cgctctggtg cttctggccg ttgtaaggga tgatggtgaa ctcccaggcg gggatcagct
     5461 tcttcaggct ggcctccagg atggtggcct tctgggtctt gtacttgaac tgcaggctct
     5521 tgttcacgat gtcgaagctc aggctgttgc tgatgatggt gttgtagctc atgaaggtgg
     5581 ccctcttgat ggcggtgccg ttgtgggtga tcatccagca caggtaggtc agctcggcgg
     5641 cacagctggc gatcttctcg ccgctgggcc tctcgaatct ctccacgaac tgccgcacca
     5701 gcaccttggg gggggtcttg cacaggatgt cgaactggct catcaccttc ctcttcttct
     5761 taggagccat ggtggccacg ccagaagcaa atgtaagcaa tagatggctc tgccctgact
     5821 tttatgccca gcccgagctc cttaaggatg aaaggcagcg tggcagttct gcaggcagtc
     5881 caaggcattg taagcaaagg aagtagcagt tggcagagac ataagtaggt gaccgaacag
     5941 gactgctttg aagagccata aacagagcca ttggcattta gggctggagg ggggtggaaa
     6001 cacaggcaaa ggaacagaag attcctg
//
