LOCUS       sequence-470510-        4744 bp    DNA     circular SYN 01-MAY-2025
DEFINITION  AiE0693h 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 4744)
  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 4744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 1, 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: AiP12695 - pAAV-AiE0693h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      266..275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(272..274,275..277,278..988)
                     /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(317..338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(578..597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     925..946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1081..1101)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1288..1399
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1517..1657
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1517..1646
                     /label=AAV2 ITR
     rep_origin      1732..2187
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1819..1838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2029..2050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2204..2223)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2323..2345
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2383..2401)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2469..2573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2574..2642,2643..3434)
                     /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(2792..2811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3605..4193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4094..4113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4255..4384
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgggcctgtg gaaacaggga gaagaatctc ttatccagca ggtctgaggg ccacaggaac
       61 ataagtaaca ctgaagagtg aggctgatcc ctggggtttg cgcgagcctg caaatgcatg
      121 attcaacagc actcgcctgg tgcctctggg aggcaagctg gatgaggatg ttgagctcgg
      181 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
      241 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
      301 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      361 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
      421 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
      481 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      541 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      601 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      661 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      721 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
      781 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      841 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
      901 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      961 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     1021 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1081 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1141 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     1201 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     1261 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     1321 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1381 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1441 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     1501 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     1561 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     1621 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     1681 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     1741 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1801 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1861 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1921 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     1981 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2041 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     2101 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2161 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2221 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2281 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2341 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2401 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2461 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2521 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2581 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2641 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2701 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2761 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2821 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2881 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2941 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3001 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3061 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3121 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3181 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3241 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3301 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3361 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3421 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca
     4261 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc
     4321 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     4381 tcctgcggcc gcacgcgtag aaagcaagag caaagggcag gtcttccagg agagacagac
     4441 agacaaagaa aaagcaaggg aggtggagag tgtgtgagag agggagtgag aaagggctca
     4501 gccatttgtc tgggcaggtc tgcggaactc aaatccggct catctgcatc tcctcaaata
     4561 aacttctggg aagtgaatgc ctgaagcagg cagagtgcag agaagtcagc aaatatgtgc
     4621 gtagtgaatt ccaagccttc tgttcccagc tcctgggagg ggccgcagtg ccttggggtg
     4681 tcctgccgtg tctccatcgg gtcccctgcc tggctgctct aaccctgagg aatggcctcc
     4741 attg
//
