LOCUS       sequence-460626-        5752 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE1404m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  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 5752)
  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: AiP15028 -
            pAAV-AiE1404m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5752
                     /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      1105..1114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1111..1113,1114..1116,1117..1827)
                     /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(1417..1436)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1764..1785
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1837..1893
                     /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             1906..1971
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2002..2379
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2466..2486)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2673..2784
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2902..3042
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2902..3031
                     /label=AAV2 ITR
     misc_feature    3049..3076
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3077..3104
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3173..3628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3260..3279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3470..3491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3645..3664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3764..3786
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3824..3842)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3910..4014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4015..4083,4084..4875)
                     /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(4233..4252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5046..5634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5535..5554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5691..5718
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5719..5746
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgtat aagatgacta ctggcctata tttacctgac
      181 aagaatctat tcctccttct tcatatcctg cacatatcat actttcagtg atgttgtatt
      241 ctggcagctg ttgctggcac ttttcattgg atataagagg aacatcagct tctttcagca
      301 catcaacagt agaacctgct tgaaatggga aatgcagaca gccagtcagt aatgattaac
      361 aggcatcctc gaatgaaacg ccatccatct gatttatccc agcattttgc tctcattatc
      421 aagtactatg tggatgtata atggaactaa ctaccaacct tgcactttca agtacataat
      481 aaacccatct gtgcactttg gtaaatggag agcatttctt tcttcctttc ctaaggggta
      541 gtaagaattg tggattggac tagggtggtc ttttaggcat tctcagtatt ttttaaagag
      601 ttacagtgtt gctggtagaa aggaagtctg ccagcacaaa gtgttcattc atggaaatca
      661 gcacagattt ggatatccag aagagctttt catccagcac ctgaggaggt gctccctcct
      721 gtataaaggc attcagctcc tttgtggttc tgtgtgtgcg agagcaggtg cacgctgaac
      781 aggggacaaa gccagctgaa ctgtgctcta ctcttggggc aatatgcttg ccatcccagg
      841 caggtgaaga gaaaggaagc atatttcagc attgtgtgtt tgcatttgtt ttatcacaaa
      901 tcactcactg caaggatgaa cctctcctta ctaaactact gttcccaaga agcttgtgaa
      961 gcaagcaggt gttagtacct tcagaaaagg aacataagca tttattcatg agagctcggg
     1021 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
     1081 ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1321 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1561 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1621 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa
     1861 caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat
     1921 gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc
     1981 agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     2041 tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc
     2101 aaggagagct attccatcta tgtgtataag gttctgaagc aggtccaccc tgacaccggc
     2161 atttcgtcca aggccatggg catcatgaac tcgtttgtga acgacatttt cgagcgcatc
     2221 gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag
     2281 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     2341 ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga
     2401 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2461 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2521 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2581 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2641 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2701 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2761 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2821 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2881 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2941 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3001 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgccta tcagcacaca
     3061 attgcccatt atacgcgcgt ataatggact attgtgtgct gatatgatgc ggtattttct
     3121 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3181 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3241 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3301 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3361 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3421 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3481 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3541 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3601 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3661 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3721 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3781 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3841 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3901 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5161 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5641 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca tatcagcaca
     5701 caattgccca ttatacgcgc gtataatgga ctattgtgtg ctgataacat gt
//
