LOCUS       sequence-432558-        5009 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2428m 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 5009)
  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 5009)
  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: AiP1746 - pAAV-AiE2428m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(293..312)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     412..434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(472..490)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        558..662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(663..731,732..1523)
                     /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(881..900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1694..2282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2183..2202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2344..2473
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3364..3373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3370..3372,3373..3375,3376..4086)
                     /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(3415..3436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3676..3695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4023..4044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4179..4199)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4386..4497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4615..4755
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4615..4744
                     /label=AAV2 ITR
     rep_origin      join(4830..5009,1..276)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4917..4936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
       61 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      121 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
      181 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
      241 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
      301 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
      361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
      481 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      661 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      721 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      781 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      841 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      901 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      961 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1021 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1081 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1141 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1201 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1261 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1321 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1381 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1441 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1501 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1561 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1621 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1681 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1741 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1801 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     1861 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1921 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1981 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2041 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2101 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2161 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2221 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2281 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2341 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     2401 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2461 actaggggtt cctgcggccg cacgcgttgt aagagtaata atcgctcatt tattttcctg
     2521 taaatgcttc tattgctaac caggccttgg ccatttttcc catgggaaat ggagttgttg
     2581 gcttcccctg gttcctgttc accatccagg aagtcgaata aataatgacc gtagcccata
     2641 ctggtgggat ggatacagga ggaggactga aagcaggaga atatcaaacc catcaaaaat
     2701 caccgagatc agtgcaaaga ctgaagcaga aagacacagc ctctaaatag gagcggcctt
     2761 aatgaaagtg catcacctca gagccaacat gaagtctgca gatgggatgc acctgcctcc
     2821 tgagaacctc acagagggaa atcaagcatc agaacccaga atggaagcag aaaaccccca
     2881 atcagactcc tcggagacac ttcagaaaag gaactctcag cttgcaaaac aacaggtgca
     2941 tctctaaaga gatgtggctc ctgcccccac cccataccac cgcaccctga gattgcagca
     3001 ctgctctgaa ctctccaatc aagcctcttt ctttggtcac tcttaaagtg gttggtttcc
     3061 agcactaacc ctcactcctc caccgaaatg aaatctgcta gctttggctg atgtttaaac
     3121 agcttcaatc tctggggtct tttgtgagag aggataattt taacactggt gataaaccca
     3181 ccagaatttc caggtgagag gagaagccta gtggagcctt tatccagctg ttgaagacag
     3241 gacctgggaa ctcttgggac aaagacatcg gagctcgggc tgggcataaa agtcagggca
     3301 gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct agcgctaccg
     3361 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3421 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3481 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     3541 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
     3601 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3661 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3721 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3781 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     3841 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     3901 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3961 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     4021 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4081 tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca acctctggat
     4141 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4201 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4261 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     4321 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc tcgagagatc
     4381 ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4441 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4501 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4561 ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac
     4621 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4681 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4741 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4801 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4861 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4921 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4981 tcgggggctc cctttagggt tccgattta
//
