LOCUS       sequence-465240-        4756 bp    DNA     circular SYN 07-JUL-2025
DEFINITION  AiE0486m 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 4756)
  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 4756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 7, 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: AiP1983 - pAAV-AiE0486m-minBG-iCre-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4756
                     /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      454..463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             466..486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1583..1603)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1790..1901
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2019..2159
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2019..2148
                     /label=AAV2 ITR
     rep_origin      2234..2689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2321..2340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2531..2552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2706..2725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2825..2847
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2885..2903)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2971..3075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3076..3144,3145..3936)
                     /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(3294..3313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4107..4695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4596..4615
                     /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 gcgtaaaatc aaatcagttg caaaatccat acagtcttcc
      181 agtttctcag ctgtggaaac tttgtgtctc ttcttttcca ccaaaacagc tatctcgtct
      241 ttcaagtcct tgctgaaaaa aaaaagtgtt aaatttttct caatttagac ttccagcaaa
      301 acagattctg cttgttgaaa ggcattggtg ttcatgagta agtgagtttc cattttattt
      361 cttcacaaat aagttacatg aagtacttga gctcgggctg ggcataaaag tcagggcaga
      421 gccatctatt gcttacattt gcttctggga tccgccacca tggtgcccaa gaagaagagg
      481 aaagtctcca acctgctgac tgtgcaccaa aacctgcctg ccctccctgt ggatgccacc
      541 tctgatgaag tcaggaagaa cctgatggac atgttcaggg acaggcaggc cttctctgaa
      601 cacacctgga agatgctcct gtctgtgtgc agatcctggg ctgcctggtg caagctgaac
      661 aacaggaaat ggttccctgc tgaacctgag gatgtgaggg actacctcct gtacctgcaa
      721 gccagaggcc tggctgtgaa gaccatccaa cagcacctgg gccagctcaa catgctgcac
      781 aggagatctg gcctgcctcg cccttctgac tccaatgctg tgtccctggt gatgaggaga
      841 atcagaaagg agaatgtgga tgctggggag agagccaagc aggccctggc ctttgaacgc
      901 actgactttg accaagtcag atccctgatg gagaactctg acagatgcca ggacatcagg
      961 aacctggcct tcctgggcat tgcctacaac accctgctgc gcattgccga aattgccaga
     1021 atcagagtga aggacatctc ccgcaccgat ggtgggagaa tgctgatcca cattggcagg
     1081 accaagaccc tggtgtccac agctggtgtg gagaaggccc tgtccctggg ggttaccaag
     1141 ctggtggaga gatggatctc tgtgtctggt gtggctgatg accccaacaa ctacctgttc
     1201 tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca cctcccaact gtccacccgg
     1261 gccctggaag ggatctttga ggccacccac cgcctgatct atggtgccaa ggatgactct
     1321 gggcagagat acctggcctg gtctggccac tctgccagag tgggtgctgc cagggacatg
     1381 gccagggctg gtgtgtccat ccctgaaatc atgcaggctg gtggctggac caatgtgaac
     1441 attgtgatga actacatcag aaacctggac tctgagactg gggccatggt gaggctgctc
     1501 gaggatgggg actaagaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa
     1561 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1621 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1681 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1741 gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct
     1801 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1861 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1921 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1981 agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     2041 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     2101 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     2161 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     2221 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2281 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     2581 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     2641 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     2701 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2761 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2821 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2881 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2941 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3001 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3061 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3121 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3181 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3241 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3301 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3361 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3421 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3481 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3541 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3601 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3661 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3721 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3781 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3841 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3901 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3961 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4021 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4081 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4141 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4201 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4261 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4321 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4381 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4441 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4501 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4561 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4621 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4681 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4741 ccttttgctc acatgt
//
