LOCUS       sequence-325770-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, 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 JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4692)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP11258 - pAAV-eHGT_022h-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..27
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11..27
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(744..746,747..749,750..1460)
                     /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(789..810)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1050..1069)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1397..1418
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1553..1573)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1753..1864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2103..2120)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2103..2119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2112..2134)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2328..2347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2447..2469
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2507..2525)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2593..2697
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2698..2766,2767..3558)
                     /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(2916..2935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3729..4317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4218..4237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4605..4626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4641..4646,4647..4664,4665..4671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(4679..4692,1..3)
                     /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     join(4684..4692,1..14)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caatttcaca caggaaacag ctatgaccat gattacgcca agctctcgag atctagaaag
       61 cttcccgggg ggatctgggc cactccctct ctgcgcgctc gctcgctcac tgaggccgcc
      121 cgggcaaagc ccgggcggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      181 ccatcactag gggttcctgg aggggtggag tcgtgaccta ggacgcgtag tcctgtggat
      241 accatctgag cctgtgccct tttctctcta ctcttgagca agtgcccatc aggagccaca
      301 tttgagtcag gacctgccaa gagcattttc atctcatcct cagttcccct tttgtctgga
      361 accactcgct atgagtttct atgagatcgt agccaggttt gaccaaacca ctcctgcaca
      421 catgttgttt tgaaagatta gcacagaatc caacacaaca tttcccttgg attcatcttt
      481 caagccaaca gagaaagagc tatttccaaa aaaataacac attattaacc aatgaataag
      541 agccaagaaa acaatgtagc cagcatattt attccaaaca gcttgtttag aatgcaaatt
      601 gcaaatacat ctatttccac ccggctcttt ggggaaccat gctggagctc gggctgggca
      661 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
      721 agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      781 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      841 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
      901 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
      961 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1021 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1081 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1141 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1201 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1261 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1321 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     1381 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1441 gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccgcgaatt cgatatcata
     1501 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     1561 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1621 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     1681 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     1741 tggctcgagc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1801 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1861 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1921 agggggagga ttgggaagac aatagcaggc atgactagtc cactccctct ctgcgcgctc
     1981 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2041 cctcagtgag cgagcgagcg cgcagagagg gacagatccg ggcccgcatg cgtcgacaat
     2101 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2161 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2221 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2281 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     2341 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2401 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2461 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2521 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2581 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2641 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2701 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2761 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2821 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2881 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2941 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3001 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3061 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3121 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3181 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3241 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3301 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3361 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3421 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3481 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3541 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3601 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3661 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3721 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3781 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3841 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3901 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3961 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4021 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4081 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4141 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4201 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4261 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4321 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4381 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4441 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4501 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4561 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4621 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4681 gtgagcggat aa
//
