LOCUS       sequence-462678-        4868 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0019m 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 4868)
  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 4868)
  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: AiP11276 - scAAV-AiE0019m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      858..867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(864..866,867..869,870..1580)
                     /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(909..930)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1170..1189)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1517..1538
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1673..1693)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1873..1984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2223..2240)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2223..2239)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2232..2254)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2448..2467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2567..2589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2627..2645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2713..2817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2818..2886,2887..3678)
                     /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(3036..3055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3849..4437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4338..4357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4725..4746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4761..4766,4767..4784,4785..4791)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4799..4815
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4804..4826
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4823..4839
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4823..4839
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctaggacgc gtttctccat
      181 ccatgtcctc ctgacttccc tctgctctgt caactccctc cataagcacc actaaaccgc
      241 ggagataatc actcctccga ggccggcagt tctcccacat aacctgaagg cactgccatg
      301 ggtttgtcac aacagatcat ctcttgaact ctgacagcca gccctaggag ctggaagagt
      361 gctcattttc ctcaccctgc cagagtttca gaaaggactc agctcaaaat agatgctatt
      421 gaagctgaga cacacacaga ggcacacagc ctatcaacgt ctcctcagtg gcccctcctc
      481 accctccaac cctaccgagt acgtgagagc cgccgctgga agcttcgtaa tccaagacca
      541 ttaagtattc atggggctgg caaccacaaa tagaaactga gttggctcat tccttttcaa
      601 attagtataa tgacattatc tgggaaatga tgatttttaa aaggagttca gaatattggt
      661 ttgaatttgc ttcctactct tctgctcagg cagttccctc tgtctttccc aggtgtggtg
      721 tggagaccct cgtgttccgt ggcaggttcc caaggagaca cacggagctc gggctgggca
      781 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
      841 agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      901 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      961 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
     1021 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
     1081 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1141 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1201 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1261 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1321 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1381 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1441 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     1501 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1561 gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata
     1621 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     1681 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1741 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     1801 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1861 tggctcgagc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1921 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1981 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2041 agggggagga ttgggaagac aatagcaggc atgactagtc cactccctct ctgcgcgctc
     2101 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2161 cctcagtgag cgagcgagcg cgcagagagg gacagatccg ggcccgcatg cgtcgacaat
     2221 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2281 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2341 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2401 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     2461 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2641 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2701 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2761 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2821 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2881 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2941 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3001 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3061 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3121 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3181 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3241 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3301 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3361 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3421 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3481 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3541 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3601 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3661 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3721 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3781 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3841 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3901 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3961 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4021 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4081 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4141 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4201 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4261 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4321 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4381 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4441 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4501 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4561 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4621 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4681 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4741 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4801 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctcg
     4861 agatctag
//
