LOCUS       sequence-462664-        4981 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0644h 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 4981)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP12683 - pAAV-AiE0644h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4981
                     /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      993..1002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(999..1001,1002..1004,1005..1715)
                     /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(1044..1065)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1305..1324)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1652..1673
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1808..1828)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2015..2126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2244..2384
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2244..2373
                     /label=AAV2 ITR
     rep_origin      2459..2914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2546..2565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2756..2777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2931..2950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3050..3072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3110..3128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3196..3300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3301..3369,3370..4161)
                     /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(3519..3538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4332..4920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4821..4840
                     /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 gcgttgccag ccacttctac cttcaagggg gactcgcggc
      181 cgggtttacg cgcgaaggat gggtgccagg gaaactgact tctagacgac ccgctccgcg
      241 agagactgac aactcagtcc tgcgggcttt cggggcgccc agaccagtcc ttagccagag
      301 gatctggggc tgcacagctc ccggaccctg gagggggtgg tccgggggtg ggagactgcg
      361 agcatcagcc cacgccggcc ccctcccacc ctgcggcggc gaggtgggaa gggccaggat
      421 gcgagcttag cacctcccct agaaatgcag cacttcgccc ccccacctcc cctgccctcg
      481 gcgctccctc tttcacgcgc tccctcccgc ctccttccca agggcgccct tcttctgccc
      541 ccagctcacg tctgaatccc tcggcgcccc ctttctcttc tcctagcccc ttcctcacgt
      601 cccctgcctc cgggtatttc ctctctccaa tcccccaccc ccgcaccgcc tgattccgag
      661 gggcgggagc gcattgggct gcgcacgggt gggggcgccg cgccagcttc gcgtagctgc
      721 tctgacgccg ctgccgccgc cgccgccgcc gccgccctcc gcagcccagc tcgcgccccg
      781 cggcagctcc gcagtgcact agccaccacc gccgccgccg ccgctccgcc agacctgctg
      841 ccagcttgcc cggtccagcc ctgagagagc ctcgaacgcc agctgcgagg gtcatgagcg
      901 agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg
      961 atccagatct ttcgaagcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
     1141 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
     1201 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1261 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1321 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1381 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1441 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1501 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1561 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1621 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1681 ccgggatcac tctcggcatg gacgagctgt acaagtaagt cgacggcgcg ccgcggccgc
     1741 gaattcgata tcataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     1921 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     1981 gcactgacaa ttccgtggct cgagagatct tcgactgtgc cttctagttg ccagccatct
     2041 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2101 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2161 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgagatc
     2221 tcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2281 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2341 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     2401 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2461 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2521 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2581 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2641 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2701 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2761 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2821 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2881 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     2941 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3001 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3061 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3121 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3181 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3241 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3301 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3361 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3421 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3481 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3541 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3601 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3661 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3721 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3781 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3841 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3901 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3961 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4021 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4081 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4141 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4201 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4261 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4321 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4381 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4441 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4501 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4561 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4621 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4681 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4741 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4801 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4861 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4921 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4981 t
//
