LOCUS       sequence-459706-        6721 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 6721)
  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 6721)
  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.RMacL3-03.EGFP.BCA12.WPRE
FEATURES             Location/Qualifiers
     source          1..6721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..34)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     225..246
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        409..513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(514..582,583..1374)
                     /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(732..751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1545..2133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2034..2053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2421..2442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2457..2462,2463..2480,2481..2487)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2495..2511
                     /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     2500..2522
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2519..2535
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2519..2535
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2573..2702
                     /label=AAV2 ITR
     regulatory      4171..4180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4177..4179,4180..4182,4183..4896)
                     /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(4177..4179,4180..4182,4183..4893)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4222..4243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4483..4502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4830..4851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5503..5523)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5921..5932)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6053..6174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6064..6083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6118..6137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6343..6472
                     /label=AAV2 ITR
     primer_bind     complement(6491..6508)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6491..6507)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6500..6522)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(6649..6721,1..383)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
       61 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      121 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
      181 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
      241 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
      301 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      361 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
      421 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      481 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      541 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      601 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      661 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      721 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      781 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      841 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      901 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      961 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1021 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1081 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1141 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1201 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     1261 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1321 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1381 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1441 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1501 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1561 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1621 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1681 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1741 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1801 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1861 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1921 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1981 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2041 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2101 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2161 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2221 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2281 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2341 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2401 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     2461 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     2521 ggaaacagct atgaccatga ttacgccaga tttaattaag gccttaatta ggctgcgcgc
     2581 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     2641 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     2701 cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagat
     2761 cggaattcaa cgctgtgctc gacctggtca acaacaagat gcgggtgagc cccagaaccc
     2821 ttgccagcca cgccgcgccc tccttccctc gcccttaact gtgatgaagg ggagcggcga
     2881 cttactgtct ccctgacagc gtcatggcta tggcctcgtg tctgccttgg cagcgtcatg
     2941 ctggggcttc agtttgagga cagctcacca aaagtgcact gtgaccacct gtcctttgta
     3001 caaaatgggg ttttctcaag cagtgtagta cttgggcttt ttaaatttag taagccaatc
     3061 taaggaaaaa agttatattc agtaagtgtg ggcagtcctg ccggagcttt ccccaagacc
     3121 ggctgccagc aggtgccagt cttcactcag gagaccagca ccccgagcgg tcctggcact
     3181 ccctcttgcc gttccctcct tagtccagat cagcagcttc tctgtggccc ccagagcatt
     3241 tagtcacagg cagtaaaccc accatgctgt cctagtttcc atgctctagc ctgcaggcag
     3301 gaacatccaa actgagcagc cggggtcccc cccacccccc accccgcccc acgcggcaac
     3361 tttgagcctg tgctgggaca gagcctctag ttcctaaatt agtccatgag gtcagaggca
     3421 gcactgccat tgtaacgcga ttggagagga tcacgtcacc ggacacgccc ccaggcatct
     3481 ccctgggtct cctaaacttg gcggggagaa gttttagccc ttaagtttta gcctttaacc
     3541 cccatattca gaactgtgcg agttggcgaa accccacaaa tcacaacaaa ctgtacacaa
     3601 caccgagcta gaggtgatct ttcttgtcca ttccacacag gccttagtaa tgcgtcgcca
     3661 tagcaacagt gtcactagta gcaccagcac ttccccacac cctccccctc aggaatccgt
     3721 actctccagt gaaccccaga aacctctgga gagttctgga caagggcgga acccacaact
     3781 ccgattactc aagggaggcg gggaagctcc accagacgcg aaactgctgg aagattcctg
     3841 gccccaaggc ctcctccggc tcgctgattg gcccagcgga gagtgggcgg ggccggtgaa
     3901 gactccttaa aggcgcaggg cggcgagcag gtcaccagac gctgacagct actcagaacc
     3961 aaatctggtt ccatccagag acaagcgaag acaagagaag cagagcgagc ggcgcgttcc
     4021 cgatcctcgg ccaggaccag ccttccccag agcatccctg ccgcggagcg caaccttccc
     4081 aggagcatcc ctgccgcgga gcgcaacttt ccccggagca tccacgccgc ggagcgcagc
     4141 cttccagaag cagagcgcgg cgccgctagc gccaccatgg tgagcaaggg cgaggagctg
     4201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     4261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     4321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     4381 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     4621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     4681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     4741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     4801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4861 gggatcactc tcggcatgga cgagctgtac aagtaagttt aaacacaggg ttaatcatta
     4921 actacaggca gtagtaacgc tcatcgtaga gcttcaacat aggacaggtg cagaggtcat
     4981 tggaacagcc ggaatcttgt attatctatg accttggacc agacgccaac acatggaatt
     5041 gcctagaaag caatcaacgc aatcaagaca cgctccaccg cttagagtat attagctgat
     5101 gtcttagcac tgtcggttaa ggatctgaac gcgagctcag agagagtgat ggttaggtca
     5161 acgtcattta tgatctccgt ccaaccttca tccacaagcg cctactgaat atataggtgg
     5221 tctctcgccg ataatagtgg aaagagtcgt aatatgcatt atgctggact agctaccgag
     5281 actaaggatg agttagtcga tcttaggact tcacaacgat tggattctca cggtcagtcg
     5341 atataaggga cacgataagc tcccagttaa tcgccattaa taccgaccat acgcgtgcag
     5401 gagatgttgc accgtcaagc ctatagatag ccaccgtgcg gccgcgatcg ctcaacctct
     5461 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5521 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5581 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5641 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5701 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5761 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5821 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5881 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5941 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6001 tcagacgagt cggatctccc tttgggccgc ctccccgcga tcgcagacat gataagatac
     6061 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     6121 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     6181 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     6241 aagtaaaacc tctacaaatg tggtagtcga aattcccgat aaggatcttc ctagagcatg
     6301 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     6361 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     6421 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     6481 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6541 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6601 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     6661 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6721 g
//
