LOCUS       sequence-466211-        6375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2010m 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 6375)
  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 6375)
  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: AiP1200 - pAAV-AiE2010m-minBG-IRES2-iCre-WPRE-...
FEATURES             Location/Qualifiers
     source          1..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             232..252
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1325..1913
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1378..1398)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1796..1807)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1945..2421
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2144..2163)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2461..2601
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2461..2590
                     /label=AAV2 ITR
     rep_origin      2676..3131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2763..2782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2973..2994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3148..3167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3267..3289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3327..3345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3413..3517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3518..3586,3587..4378)
                     /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(3736..3755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4549..5137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5038..5057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5199..5328
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    join(6008..6375,1..206)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    join(6031..6375,1..208)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6175..6192)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
       61 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
      121 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
      181 ttttcctttg aaaaacacga tgataatatg gccacagcta gcaccatggt gcccaagaag
      241 aagaggaaag tctccaacct gctgactgtg caccaaaacc tgcctgccct ccctgtggat
      301 gccacctctg atgaagtcag gaagaacctg atggacatgt tcagggacag gcaggccttc
      361 tctgaacaca cctggaagat gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag
      421 ctgaacaaca ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac
      481 ctgcaagcca gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg
      541 ctgcacagga gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg
      601 aggagaatca gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt
      661 gaacgcactg actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac
      721 atcaggaacc tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt
      781 gccagaatca gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt
      841 ggcaggacca agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt
      901 accaagctgg tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac
      961 ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc
     1021 acccgggccc tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat
     1081 gactctgggc agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg
     1141 gacatggcca gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat
     1201 gtgaacattg tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg
     1261 ctgctcgagg atggggacta atgaggcgcg ccgcggccgc gaattcgata tcaagcttat
     1321 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1381 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1441 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     1501 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     1561 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     1621 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1681 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     1741 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     1801 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     1861 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     1921 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     1981 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     2041 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     2101 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     2161 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     2221 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     2281 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     2341 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     2401 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     2461 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2521 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2581 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2641 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     2761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2821 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2881 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3001 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3121 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3301 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3361 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5161 acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg
     5221 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     5281 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc
     5341 gtatgtgtct tttactctga tcctcctgtt tttaccttcc aagtgctgga atcacagaca
     5401 tataccactg tgcatagcat cattacaatg ttatagtttt tcacactatg ccttgacttt
     5461 ttggaaaggc aaaccacctc ttggatttct ccttccttct ctatctctct ctctctctct
     5521 tcctccctcc gtccctccat ctcttcctcc ttcccatttt cttctctccc tatttggaca
     5581 caatataaaa taatttagat gaggtgagtt aaattgtgaa caaagtatgt gcctatacat
     5641 ggttgtaaat cagcttatca aagtgtaata ttagaagaat ttataaaaat gataaaattc
     5701 atactcaaag ttctgtgtaa agcaataata gctttatctc cttttagtta tcttgagtct
     5761 ttctatgact aacaactccc tcataggcat cttaaagagc agtaagcata agtagattcc
     5821 aaatgggaag ggagaagtgt gaaccatcac tttcatccag acttgtagat atatctgctg
     5881 cattttcaga aaccagaaac agacagtgtt ctttatctcc attgagtcta gtgtagcaac
     5941 agagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     6001 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     6061 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     6121 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     6181 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     6241 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     6301 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     6361 agtgccacgt tgtga
//
