LOCUS       sequence-316829-        4856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  As alternatives to this plasmid, the Allen Institute for Brain 
            Science recommends either https://www.addgene.org/164457/ or 
            https://www.addgene.org/164458/.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4856)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 4856)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP1269-scAAV-mscRE4-minBGpromoter-SYFP2-WPRE3...
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      846..855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(852..854,855..857,858..1568)
                     /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(897..918)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1158..1177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1505..1526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1661..1681)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1861..1972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2211..2228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2211..2227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2436..2455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2555..2577
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2615..2633)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2701..2805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2806..2874,2875..3666)
                     /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(3024..3043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3837..4425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4326..4345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4713..4734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4749..4754,4755..4772,4773..4779)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4787..4803
                     /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     4792..4814
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4811..4827
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4811..4827
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt
      181 gacctaggac gcgtataagc cttgggggca atcaaactat tacattgagt ccttggattt
      241 gctacaaatt acattttaaa tgcaatcatt ttataaaagc ttcaacactc acacttggaa
      301 gcgttaccct gttgaatatc actgactcac taacttgcat tgccatgcta acttgctttc
      361 agagagatct cagaacacat catcttctgc tatttcaata catgcacatt aatttcctat
      421 caacgtgtgc tgatcaggaa ctctgtaatc tggcaccggt gtttattttt attcctgtct
      481 attcctgttg gctcacgaaa agattgtttg agcaagtgtt ttatggtgag ttgtatcata
      541 tgtacattga tttaatctgc ccacattcag ttctacaagc ggagccaaaa aaatagagac
      601 aagcataatt ttcattcaac atgagcccct caatgcaagc caagtacctc atctggtgct
      661 cagctaaagc aacagcaatc tgttccaccc tggagacaca actggccaca gaaaacttag
      721 tgaaaagagg caatgctatg cacaggacaa atgagctcgg gctgggcata aaagtcaggg
      781 cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag ctagcgctac
      841 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      901 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      961 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     1021 cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc cgctaccccg
     1081 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1141 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1201 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1261 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1321 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1381 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1441 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     1501 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1561 tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat caacctctgg
     1621 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1681 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1741 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     1801 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gctcgagcga
     1861 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1921 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1981 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2041 gggaagacaa tagcaggcat gactagtcca ctccctctct gcgcgctcgc tcgctcactg
     2101 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2161 agcgagcgcg cagagaggga cagatccggg cccgcatgcg tcgacaattc actggccgtc
     2221 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2281 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2341 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     2401 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2461 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2521 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2581 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     2641 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     2701 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2761 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2821 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2881 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2941 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3001 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3061 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3121 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3181 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3241 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3301 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3361 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3421 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3481 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3541 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3601 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3661 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3721 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3781 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3841 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3901 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3961 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4321 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4381 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4441 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4501 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4561 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4621 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4681 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4741 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4801 caatttcaca caggaaacag ctatgaccat gattacgcca agctctcgag atctag
//
