LOCUS       sequence-459708-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L5 extratelencephalic projection 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.RMacL5ET-01.EGFP.BCA7.WPRE
FEATURES             Location/Qualifiers
     source          1..6721
                     /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      4555..4564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4561..4563,4564..4566,4567..5280)
                     /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(4561..4563,4564..4566,4567..5277)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4606..4627)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4867..4886)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5214..5235
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5887..5907)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6305..6316)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6437..6558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6448..6467)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6502..6521
                     /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 tctgtgcctg ccactcaaat agaaatcgtg
     3181 ctcaccaagt gttagggccg gttctcagta cacctactgc gtggatttgc agagggtgga
     3241 gtgcatggag ggctcctcct gggagcctgt ggccgaggct gcacagcatc tttggttgct
     3301 gttcctgtct ccaccgtgaa gatggagcca cgtccctcgc ggcccctggc tgggccccac
     3361 ctcccggaga ctcaggaagg gcttggaatt ggtgccagct tcgccggctg tgggccccac
     3421 ggcggtgatg aataacttgt ttatctgctg gagctgctcc caccgctctc tcggtgcctg
     3481 gcaggcagag gtctggaagg ggcgggcagg gacggggagg gctggctcac caggcagcca
     3541 ccgcccctcc tccctcctcc ctctgcccca ctccccaatt cagcatcaac ctccacctaa
     3601 acctgcttcc tgcgctttcc tgctcctcat ctttccccaa atcctgtttt ggatcctagt
     3661 ttccatgctc tagcctgcag gcaggaacat ccaaactgag cagccggggt cccccccacc
     3721 ccccaccccg ccccacgcgg caactttgag cctgtgctgg gacagagcct ctagttccta
     3781 aattagtcca tgaggtcaga ggcagcactg ccattgtaac gcgattggag aggatcacgt
     3841 caccggacac gcccccaggc atctccctgg gtctcctaaa cttggcgggg agaagtttta
     3901 gcccttaagt tttagccttt aacccccata ttcagaactg tgcgagttgg cgaaacccca
     3961 caaatcacaa caaactgtac acaacaccga gctagaggtg atctttcttg tccattccac
     4021 acaggcctta gtaatgcgtc gccatagcaa cagtgtcact agtagcacca gcacttcccc
     4081 acaccctccc cctcaggaat ccgtactctc cagtgaaccc cagaaacctc tggagagttc
     4141 tggacaaggg cggaacccac aactccgatt actcaaggga ggcggggaag ctccaccaga
     4201 cgcgaaactg ctggaagatt cctggcccca aggcctcctc cggctcgctg attggcccag
     4261 cggagagtgg gcggggccgg tgaagactcc ttaaaggcgc agggcggcga gcaggtcacc
     4321 agacgctgac agctactcag aaccaaatct ggttccatcc agagacaagc gaagacaaga
     4381 gaagcagagc gagcggcgcg ttcccgatcc tcggccagga ccagccttcc ccagagcatc
     4441 cctgccgcgg agcgcaacct tcccaggagc atccctgccg cggagcgcaa ctttccccgg
     4501 agcatccacg ccgcggagcg cagccttcca gaagcagagc gcggcgccgc tagcgccacc
     4561 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4621 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4681 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4741 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4801 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4861 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4921 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4981 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     5041 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5101 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5161 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5221 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     5281 gtttaaacac agggttaatc attaactaca gtcttctcgt ccttactaat tgctcgtgcc
     5341 acaaggaaca tcgacctatg tggtacgatg gtatcgagaa ctacagagag ctgattgttc
     5401 gcggtgtcct gagtaccatt acgaagcggt tagtgcaatc tcaatgagac aagtggataa
     5461 ccaagtatta gccatgccta gagcagatcc tcctctagca caatggttgc gaatgagtat
     5521 ccaacatagt ccagccgtaa gttcacggac aaacatggac gacttaatca cctattcgcg
     5581 tccacatatc cgaattgttc tggtaggtaa gcctgcgagt tgatgcagga acttgtatgc
     5641 aatcttgtga cgtaacgcat gtcaacgatt agtgtatgcc gaggaaatcc gtgacttact
     5701 tcgccggata accatgccat ctactgctga actagtggat ataagcaaca tacatcgcat
     5761 caccagtaag tagtctcttc caatcgctta tgaagtatcc gttgcatcta ctctacctcg
     5821 cgcggccgcg atcgctcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5881 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5941 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6001 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6061 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6121 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6181 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6241 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6301 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6361 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6421 gcgatcgcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     6481 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     6541 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     6601 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtag tcgaaattcc
     6661 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     6721 a
//
