LOCUS       sequence-470739-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0711h 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 5114)
  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 5114)
  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: AiP13660 -
            pAAV-AiE0711h-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   291..420
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(482..1070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(562..581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1241..2032,2033..2101))
                     /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     1864..1883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2102..2206)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2274..2292
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2330..2352)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2452..2471
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2488..2943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2625..2646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2837..2856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3018..3158
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3029..3158
                     /label=AAV2 ITR
     polyA_signal    3276..3387
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3574..3594
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3681..4058)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(4089..4154)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4167..4223)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(4233..4943,4944..4946,4947..4949))
                     /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(4275..4296)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4624..4643
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      4946..4955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttggcagaaa gccatcttcc ccttcaggaa tatagaacac agctgtagga attgcatgaa
       61 aggatgctga ttttgaagga tctgaaaatt gcgatggctg cctaggagca tcgtaaacca
      121 cccaggccca caaggaagga aacacagtca taacacttca tggatgccta gagccaagta
      181 tcagtgtgat tgtattcaag actggtgcag ccttcaaaca ttcagattac cgtcatcata
      241 aatcagcaaa gatagtgcaa caagatgaaa tccacaacgc gtgcggccgc aggaacccct
      301 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
      361 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
      421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2221 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2281 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2341 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2401 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2461 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
     2521 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
     2581 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
     2641 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2701 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     2761 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2821 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
     2881 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2941 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3001 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     3061 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     3121 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
     3181 agatctcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     3241 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     3301 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     3361 cagatggctg gcaactagaa ggcacagtcg aagatctctc gagccacgga attgtcagtg
     3421 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     3481 gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     3541 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     3601 agaataccag tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt
     3661 cgcggccgcg gcgcgcctca ttacttagcg ctggtgtact tggtgatggc cttagtaccc
     3721 tcggacacgg cgtgcttggc caactcccca ggcagcagca ggcgcacggc cgtctggatc
     3781 tcccgggagg tgatggtcga gcgcttgttg taatgcgcca ggcgggaagc ctctcctgcg
     3841 atgcgctcga aaatgtcgtt cacaaacgaa ttcatgatgc ccatggcctt ggacgaaatg
     3901 ccggtgtcag ggtggacctg cttcagaacc ttgtacacat agatggaata gctctccttg
     3961 cggctgcgct tgcgcttctt gccgcctttc ttctgcgcct tagtcaccgc cttcttggag
     4021 ccctttttcg gggcgggagc agacttcgct ggctctgggc ctccggagcc tccagatctg
     4081 agtccggact tgtcatcgtc atccttgtaa tcgatgtcat gatctttata atctccgtca
     4141 tggtctttgt agtctccact agcaccaggc cctggattct cctctacatc ccctgcctgt
     4201 ttcaacagtg agaagttggt agcaccagag cccttgtaca gctcgtccat gccgagagtg
     4261 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     4321 ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     4381 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
     4441 tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     4501 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     4561 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     4621 cgggtcttgt agttgccgtc gtctttgaag aagatggtgc gctcctggac gtagccttcg
     4681 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     4741 tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     4801 gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     4861 acgctgaact tgtggccgtt gacatcgccg tccagctcga ccaggatggg caccaccccg
     4921 gtgaacagct cctcgccctt ggagaccatg gtggcgaccg gtagcgctag cttcgaaaga
     4981 tctggatccc agaagcaaat gtaagcaata gatggctctg ccctgacttt tatgcccagc
     5041 ccgagctcct ttctgagact gtaagatcta cagttattta atgtctcaag gttgattgaa
     5101 atggctataa ttcc
//
