LOCUS       sequence-431679-        4561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EYFP from pCALM1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4561)
  AUTHORS   Wang J, Lin J, Chen Y, Liu J, Zheng Q, Deng M, Wang R, Zhang Y, Feng
            S, Xu Z, Ye W, Hu Y, Duan J, Lin Y, Dai J, Chen Y, Li Y, Luo T, Chen
            Q, Lu Z
  TITLE     An ultra-compact promoter drives widespread neuronal expression in 
            mouse and monkey brains.
  JOURNAL   Cell Rep. 2023 Nov 28;42(11):113348. doi: 
            10.1016/j.celrep.2023.113348. Epub 2023 Oct 31.
  PUBMED    37910509
REFERENCE   2  (bases 1 to 4561)
  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 Rep.";
            date: "2023-11-28"; volume: "42(11):113348. doi"; pages: " 
            10.1016/j.celrep.2023.113348. Epub 2023 Oct 31"
COMMENT     Sequence Label: pAAV-pCALM1-EYFP-WPRE-bGHpA
FEATURES             Location/Qualifiers
     source          1..4561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      272..281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(278..280,281..283,284..997)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(323..344)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(584..603)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     931..952
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1009..1597
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1062..1082)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1480..1491)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   1824..1964
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1824..1953
                     /label=AAV2 ITR
     rep_origin      2039..2494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2126..2145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2336..2357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2511..2530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2630..2652
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2690..2708)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2776..2880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2881..2949,2950..3741)
                     /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(3099..3118)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3912..4500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4401..4420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactggatc cgccaccatg gtgagcaagg gcgaggagct
      301 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
      361 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
      421 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
      481 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
      541 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
      601 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
      661 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
      721 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
      781 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
      841 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
      901 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      961 cgggatcact ctcggcatgg acgagctgta caagtaagtc gactcgataa tcaacctctg
     1021 gattacaaaa tttgtgaaag atcgactggt attcttaact atgttgctcc ttttacgcta
     1081 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1141 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1201 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     1261 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     1321 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1381 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     1441 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     1501 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     1561 cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggagctctgc ccctcccccg
     1621 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1681 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1741 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatatcttt caccggtgtg
     1801 gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     1861 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     1921 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     1981 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     2041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2101 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2221 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     2281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2341 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     2401 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2461 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     2521 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     2581 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     2641 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     2701 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2821 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2881 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2941 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3001 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3061 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3121 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3181 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3241 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3301 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3361 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3421 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3481 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3541 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3601 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3661 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3721 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3781 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3841 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4021 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4081 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4141 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4201 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4261 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4321 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4381 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4441 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4501 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4561 t
//
