LOCUS       sequence-461923-        5117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0050m_3xC2 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 5117)
  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 5117)
  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: AiP13775 - pAAV-AiE0050m_3xC2-minBG-iCre-BGHpA...
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1062..1082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2151..2262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2380..2520
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2380..2509
                     /label=AAV2 ITR
     rep_origin      2595..3050
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2682..2701)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2892..2913
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3067..3086)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3186..3208
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3246..3264)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3332..3436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3437..3505,3506..4297)
                     /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(3655..3674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4468..5056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4957..4976
                     /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 gcgtaggcaa cttgttaaaa atctcagtca aacttacata
      181 ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag tttcttattc
      241 agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa agtcacaaat
      301 acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact tttattttgc
      361 ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca gcaggaaata
      421 ggcaacttgt taaaaatctc agtcaaactt acatactata tagaacagca tggtgaattt
      481 agggcacatg gatataaaat ggaagtttct tattcagtag cagcaacttg tgggcacagg
      541 agttggcaaa gataaaaatg tccaaagtca caaatacaat gtatagttag tcataggtgc
      601 tgttatttgc ctcaaaaaat agacttttat tttgcctttc ttttctttaa ccacactcaa
      661 aattagagaa cagagacaaa acccagcagg aaataggcaa cttgttaaaa atctcagtca
      721 aacttacata ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag
      781 tttcttattc agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa
      841 agtcacaaat acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact
      901 tttattttgc ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca
      961 gcaggaaatg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1021 tgcttctggg atccagatct ttcgaagcta gcaccatggt gcccaagaag aagaggaaag
     1081 tctccaacct gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg
     1141 atgaagtcag gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca
     1201 cctggaagat gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca
     1261 ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca
     1321 gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga
     1381 gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca
     1441 gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg
     1501 actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc
     1561 tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca
     1621 gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca
     1681 agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg
     1741 tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc
     1801 gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc
     1861 tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc
     1921 agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca
     1981 gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg
     2041 tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg
     2101 atggggacta atgaggcgcg ccgaattcaa gcttctcgag agatcttcga ctgtgccttc
     2161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2341 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     2401 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2461 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     2521 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2581 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2641 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2701 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2761 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2821 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2881 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2941 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     3001 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     3061 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3121 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3181 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3241 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3301 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3361 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3421 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3481 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3541 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3601 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3661 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3721 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3781 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3841 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3901 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3961 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4021 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4081 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4141 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4201 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4261 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5101 gccttttgct cacatgt
//
