LOCUS       sequence-465228-        5225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2135m_3xC2 is an optimized enhancer sequence, designed to induce 
            cre-dependent recombination in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  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 5225)
  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: AiP1536 - pAAV-AiE2135m_3xC2-minBG-iCre-HGHpA
FEATURES             Location/Qualifiers
     source          1..5225
                     /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      891..900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             903..923
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1972..2448
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2171..2190)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2488..2628
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2488..2617
                     /label=AAV2 ITR
     rep_origin      2703..3158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2790..2809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3000..3021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3175..3194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3294..3316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3354..3372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3440..3544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3545..3613,3614..4405)
                     /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(3763..3782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4576..5164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5065..5084
                     /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 gcgttccaag gccccctttc ctgccaaaca aagcccccag
      181 ccacccgact gccgcccaac aggcacctgg ggcctctggg cctgggaatc aaagcctgtt
      241 ttgcaaagga ggcccagtgt ttggagaaca cccatcccct gctgcccggt ccttcaaaga
      301 accggcattg ttctgcgctg agaaggcgcc tcttgacgcg ctgggcaggg tcggcttgac
      361 gctggtggac ctccaaggcc ccctttcctg ccaaacaaag cccccagcca cccgactgcc
      421 gcccaacagg cacctggggc ctctgggcct gggaatcaaa gcctgttttg caaaggaggc
      481 ccagtgtttg gagaacaccc atcccctgct gcccggtcct tcaaagaacc ggcattgttc
      541 tgcgctgaga aggcgcctct tgacgcgctg ggcagggtcg gcttgacgct ggtggacctc
      601 caaggccccc tttcctgcca aacaaagccc ccagccaccc gactgccgcc caacaggcac
      661 ctggggcctc tgggcctggg aatcaaagcc tgttttgcaa aggaggccca gtgtttggag
      721 aacacccatc ccctgctgcc cggtccttca aagaaccggc attgttctgc gctgagaagg
      781 cgcctcttga cgcgctgggc agggtcggct tgacgctggt ggaccgagct cgggctgggc
      841 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc gccaccatgg
      901 tgcccaagaa gaagaggaaa gtctccaacc tgctgactgt gcaccaaaac ctgcctgccc
      961 tccctgtgga tgccacctct gatgaagtca ggaagaacct gatggacatg ttcagggaca
     1021 ggcaggcctt ctctgaacac acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg
     1081 cctggtgcaa gctgaacaac aggaaatggt tccctgctga acctgaggat gtgagggact
     1141 acctcctgta cctgcaagcc agaggcctgg ctgtgaagac catccaacag cacctgggcc
     1201 agctcaacat gctgcacagg agatctggcc tgcctcgccc ttctgactcc aatgctgtgt
     1261 ccctggtgat gaggagaatc agaaaggaga atgtggatgc tggggagaga gccaagcagg
     1321 ccctggcctt tgaacgcact gactttgacc aagtcagatc cctgatggag aactctgaca
     1381 gatgccagga catcaggaac ctggccttcc tgggcattgc ctacaacacc ctgctgcgca
     1441 ttgccgaaat tgccagaatc agagtgaagg acatctcccg caccgatggt gggagaatgc
     1501 tgatccacat tggcaggacc aagaccctgg tgtccacagc tggtgtggag aaggccctgt
     1561 ccctgggggt taccaagctg gtggagagat ggatctctgt gtctggtgtg gctgatgacc
     1621 ccaacaacta cctgttctgc cgggtcagaa agaatggtgt ggctgcccct tctgccacct
     1681 cccaactgtc cacccgggcc ctggaaggga tctttgaggc cacccaccgc ctgatctatg
     1741 gtgccaagga tgactctggg cagagatacc tggcctggtc tggccactct gccagagtgg
     1801 gtgctgccag ggacatggcc agggctggtg tgtccatccc tgaaatcatg caggctggtg
     1861 gctggaccaa tgtgaacatt gtgatgaact acatcagaaa cctggactct gagactgggg
     1921 ccatggtgag gctgctcgag gatggggact aaggctgctc gagagatcta cgggtggcat
     1981 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2041 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2101 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2161 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2221 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2281 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     2341 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     2401 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     2461 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2521 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2581 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2641 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2701 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2761 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2881 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3001 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3061 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3121 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3181 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3361 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5221 catgt
//
