LOCUS       sequence-459705-        6722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L3 pyramidal neuron Enhancer to drive transgene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  AUTHORS   He J, Phan BN, Kerkhoff WG, Alikaya A, Hong T, Brull OR, Fredericks 
            JM, Sedorovitz M, Srinivasan C, Leone MJ, Wirfel OM, Brown A, Dauby 
            S, Tittle RK, Lin MK, Hooks BM, Bostan AC, Gharbawie OA, Byrne LC, 
            Pfenning AR, Stauffer WR
  TITLE     Machine learning identification of enhancers in the rhesus macaque 
            genome.
  JOURNAL   Neuron. 2025 May 21;113(10):1548-1561.e8. doi: 
            10.1016/j.neuron.2025.04.030.
  PUBMED    40403706
REFERENCE   2  (bases 1 to 6722)
  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"; doi: 
            "10.1016/j.neuron.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1548-1561.e8"
COMMENT     Sequence Label: pAAV.RMacL3-02.EGFP.BCA7.WPRE
FEATURES             Location/Qualifiers
     source          1..6722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR
     primer_bind     complement(154..171)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(154..170)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(163..185)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      312..767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(399..418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     609..630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        793..897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(898..966,967..1758)
                     /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(1116..1135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1929..2517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2418..2437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2805..2826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2841..2846,2847..2864,2865..2871)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2879..2895
                     /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     2884..2906
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2903..2919
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2903..2919
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2957..3086
                     /label=AAV2 ITR
     regulatory      4556..4565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4562..4564,4565..4567,4568..5281)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4562..4564,4565..4567,4568..5278)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4607..4628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4868..4887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5215..5236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5888..5908)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6306..6317)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6438..6559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6449..6468)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6503..6522
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
       61 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
      121 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
      181 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      241 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      301 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      361 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      421 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      481 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      541 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      601 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      661 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      721 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      781 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      841 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      901 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      961 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1021 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1081 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1141 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1201 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1261 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1321 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1381 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1441 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1501 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1561 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1621 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1681 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1741 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1801 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1861 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1921 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1981 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2041 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2101 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2161 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2221 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2281 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2341 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2401 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2461 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2521 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2581 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2641 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2701 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     2761 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2821 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2881 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc cagatttaat
     2941 taaggcctta attaggctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     3001 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     3061 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
     3121 tacgtagcca tgctctagga agatcggaat tcttctggtg ggatattgaa gggtaggagg
     3181 gaggatgatg agggaaggca ctgggggaca gggaaggagg agagtggggg attgtgaggg
     3241 cgaagagacc aggcagacct gccagtgaag ctccttagtc cctgagccag gatttcattt
     3301 taaatacacc aagacctttt ttagcaagtc ctagacagat ccaccattca tctggacagt
     3361 gcgtttttcc ttgcagaaaa taacctatga ccagccagca gctcctggag tgtgggctga
     3421 ggaacacagt ccagtgggcc agcactcctt catcctgtga tgccagtggc tgcgaggagc
     3481 ccacattcac ggcctaaaaa tagcccccat ctctactcca cccctcctgg gccctaggct
     3541 ttccagggac tccctctcca tgttgacttg gtttgggaca gggagtgtgt ttctggaaga
     3601 atatttctag cctgagctgc tttttccttc caagtaaggc agtttgaaag cgttcctagt
     3661 ttccatgctc tagcctgcag gcaggaacat ccaaactgag cagccggggt cccccccacc
     3721 ccccnacccc gccccacgcg gcaactttga gcctgtgctg ggacagagcc tctagttcct
     3781 aaattagtcc atgaggtcag aggcagcact gccattgtaa cgcgattgga gaggatcacg
     3841 tcaccggaca cgcccccagg catctccctg ggtctcctaa acttggcggg gagaagtttt
     3901 agcccttaag ttttagcctt taacccccat attcagaact gtgcgagttg gcgaaacccc
     3961 acaaatcaca acaaactgta cacaacaccg agctagaggt gatctttctt gtccattcca
     4021 cacaggcctt agtaatgcgt cgccatagca acagtgtcac tagtagcacc agcacttccc
     4081 cacaccctcc ccctcaggaa tccgtactct ccagtgaacc ccagaaacct ctggagagtt
     4141 ctggacaagg gcggaaccca caactccgat tactcaaggg aggcggggaa gctccaccag
     4201 acgcgaaact gctggaagat tcctggcccc aaggcctcct ccggctcgct gattggccca
     4261 gcggagagtg ggcggggccg gtgaagactc cttaaaggcg cagggcggcg agcaggtcac
     4321 cagacgctga cagctactca gaaccaaatc tggttccatc cagagacaag cgaagacaag
     4381 agaagcagag cgagcggcgc gttcccgatc ctcggccagg accagccttc cccagagcat
     4441 ccctgccgcg gagcgcaacc ttcccaggag catccctgcc gcggagcgca actttccccg
     4501 gagcatccac gccgcggagc gcagccttcc agaagcagag cgcggcgccg ctagcgccac
     4561 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4621 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4681 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4741 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4801 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4861 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4921 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4981 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     5041 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     5101 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     5161 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     5221 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     5281 agtttaaaca cagggttaat cattaactac agtcttctcg tccttactaa ttgctcgtgc
     5341 cacaaggaac atcgacctat gtggtacgat ggtatcgaga actacagaga gctgattgtt
     5401 cgcggtgtcc tgagtaccat tacgaagcgg ttagtgcaat ctcaatgaga caagtggata
     5461 accaagtatt agccatgcct agagcagatc ctcctctagc acaatggttg cgaatgagta
     5521 tccaacatag tccagccgta agttcacgga caaacatgga cgacttaatc acctattcgc
     5581 gtccacatat ccgaattgtt ctggtaggta agcctgcgag ttgatgcagg aacttgtatg
     5641 caatcttgtg acgtaacgca tgtcaacgat tagtgtatgc cgaggaaatc cgtgacttac
     5701 ttcgccggat aaccatgcca tctactgctg aactagtgga tataagcaac atacatcgca
     5761 tcaccagtaa gtagtctctt ccaatcgctt atgaagtatc cgttgcatct actctacctc
     5821 gcgcggccgc gatcgctcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5881 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5941 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6001 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     6061 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6121 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     6181 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     6241 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6301 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6361 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     6421 cgcgatcgca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     6481 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     6541 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     6601 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta gtcgaaattc
     6661 ccgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     6721 aa
//
