LOCUS       sequence-462510-        4788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0671m_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 4788)
  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 4788)
  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: AiP20026 -
            pAAV-AiE0671m_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..4788
                     /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      724..733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             736..756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1822..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2051..2191
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2051..2180
                     /label=AAV2 ITR
     rep_origin      2266..2721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2353..2372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2563..2584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2738..2757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2857..2879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2917..2935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3003..3107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3108..3176,3177..3968)
                     /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(3326..3345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4139..4727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4628..4647
                     /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 gcgtaaaggt tgcactgtat ctgaaataag ctacatttcc
      181 ttcccagctc accaaaaagc aggacatatg gatgtgaaca agagatccag gacagctgaa
      241 gtacttttaa ggaacactgt aaactttcgt ctctaaaatt aatcatctct atgtatgagt
      301 atatgttcag aaaggttgca ctgtatctga aataagctac atttccttcc cagctcacca
      361 aaaagcagga catatggatg tgaacaagag atccaggaca gctgaagtac ttttaaggaa
      421 cactgtaaac tttcgtctct aaaattaatc atctctatgt atgagtatat gttcagaaag
      481 gttgcactgt atctgaaata agctacattt ccttcccagc tcaccaaaaa gcaggacata
      541 tggatgtgaa caagagatcc aggacagctg aagtactttt aaggaacact gtaaactttc
      601 gtctctaaaa ttaatcatct ctatgtatga gtatatgttc aggagctcgg gctgggcata
      661 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
      721 ctagccacca tggtgcccaa gaagaagagg aaagtctcca acctgctgac tgtgcaccaa
      781 aacctgcctg ccctccctgt ggatgccacc tctgatgaag tcaggaagaa cctgatggac
      841 atgttcaggg acaggcaggc cttctctgaa cacacctgga agatgctcct gtctgtgtgc
      901 agatcctggg ctgcctggtg caagctgaac aacaggaaat ggttccctgc tgaacctgag
      961 gatgtgaggg actacctcct gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa
     1021 cagcacctgg gccagctcaa catgctgcac aggagatctg gcctgcctcg cccttctgac
     1081 tccaatgctg tgtccctggt gatgaggaga atcagaaagg agaatgtgga tgctggggag
     1141 agagccaagc aggccctggc ctttgaacgc actgactttg accaagtcag atccctgatg
     1201 gagaactctg acagatgcca ggacatcagg aacctggcct tcctgggcat tgcctacaac
     1261 accctgctgc gcattgccga aattgccaga atcagagtga aggacatctc ccgcaccgat
     1321 ggtgggagaa tgctgatcca cattggcagg accaagaccc tggtgtccac agctggtgtg
     1381 gagaaggccc tgtccctggg ggttaccaag ctggtggaga gatggatctc tgtgtctggt
     1441 gtggctgatg accccaacaa ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc
     1501 ccttctgcca cctcccaact gtccacccgg gccctggaag ggatctttga ggccacccac
     1561 cgcctgatct atggtgccaa ggatgactct gggcagagat acctggcctg gtctggccac
     1621 tctgccagag tgggtgctgc caccgacatg gccagggctg gtgtgtccat ccctgaaatc
     1681 atgcaggctg gtggctggac caatgtgaac attgtgatga actacatcag aaacctggac
     1741 tctgagactg gggccatggt gaggctgctc gaagatgggg actgaggcgc gccgaattca
     1801 agcttctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     2041 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2101 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2161 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2221 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2281 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2341 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2401 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2461 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2521 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2581 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2641 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2701 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2761 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2821 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2881 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     2941 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3121 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3181 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3241 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3301 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3361 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3421 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3481 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3541 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3601 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3661 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3721 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3781 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3841 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3901 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3961 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4021 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4081 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4141 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4201 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4261 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4321 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4381 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4441 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4501 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4561 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4621 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4681 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4741 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
