LOCUS       sequence-471602-        5186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2617m 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 5186)
  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 5186)
  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: AiP2042 - pAAV-AiE2617m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5186
                     /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      1198..1207
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1204..1206,1207..1209,1210..1920)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1249..1270)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1510..1529)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1857..1878
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2013..2033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2220..2331
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2449..2589
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2449..2578
                     /label=AAV2 ITR
     rep_origin      2664..3119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2751..2770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2961..2982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3136..3155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3255..3277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3315..3333)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3401..3505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3506..3574,3575..4366)
                     /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(3724..3743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4537..5125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5026..5045
                     /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 gcgtccataa tgtcagtgat tagtgtctcc acgaaagctg
      181 attttgtatt gcttttctga gtaaatgact attcaggtcc cccactcccc acaatgatgc
      241 tgcctgtgta attggcagag agggacatcc tgagaatgga ggtgtgaagg tgtaacgtgt
      301 gttaattgat acttcctaat cgcttttagg tattaagtca tgatgcagga atctgcgaca
      361 gagacaataa gcaacagttc aatgaatcaa aatggaatga gcactttaag cagccaatta
      421 gatgctggca gcagggatgg gagatcaagt ggtgacacca gctctgaagt aagcacagtg
      481 gagctgctgc atctgcaaca acagcaggta agttttgttt tcctcaaagc ttcccttaaa
      541 acaaatctgc ctgtttgctt gaacctggag actgtattca tagtaaattg agcatgttgt
      601 tttaagctaa cagatgctga gagtttattg tcatttgcaa catgattgaa aggtgaggga
      661 agagaagctt cgcattagtt acgtcatagc agtgccagag tgcccttcag gtgtcaggat
      721 gcattaattg tgttcgcagg gtgatttttt ttttttttta agagtggttc tgtaaagctt
      781 tgtaacaaat attttcgagc ctctggctta ctgcttggca ttgtgctgcg gaatgtggtg
      841 ctgattttcc accagagggc cacattactt gatacttgct tatacaatga atttcttcca
      901 agtctcagct aaggtgaaaa ggaccacgag gacttttcag agcataggtg ctggctattg
      961 ctagttgcag atattaagta ttaattaatt aatctgtgat aggttatgca acctgtaaaa
     1021 agttcattta ctcacctgtg cttctgttct gagagactcg ttccactatt ttgaatgtgc
     1081 aaatgtgtgt gtgtgtcgcc ggcggagctc gggctgggca taaaagtcag ggcagagcca
     1141 tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct accggtcgcc
     1201 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1261 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1321 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1381 accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     1441 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1501 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1561 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1621 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1681 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1741 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1801 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1861 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1921 taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata atcaacctct ggattacaaa
     1981 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2041 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2101 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     2161 tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga gatcttcgac
     2221 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2281 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2341 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2401 ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag gaacccctag
     2461 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2521 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2581 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2641 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2701 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2761 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2821 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2881 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2941 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3001 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3061 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3121 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3181 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3241 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3301 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3361 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3421 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3481 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3541 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3601 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3661 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3721 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3781 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3841 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3901 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3961 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4021 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4081 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4141 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4201 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4261 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4321 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4381 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4441 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5161 cttttgctgg ccttttgctc acatgt
//
