LOCUS       sequence-484630-        8409 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  Human RIBEYE cDNA cloned inframe into a pAAV-GFP vector driven by 
            hybrid CMV-enhancer-human-beta-actin promoter (CMV-HBA) and 
            containing downstream WPRE, Ampicillin Resistance .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8409)
  AUTHORS   Kapoor R, Schwenzer N, Dresbach T, Lehnart SE, Moser T
  TITLE     Establishing synthetic ribbon-type active zones in a heterologous 
            expression system
  JOURNAL   eLife13:RP98254
REFERENCE   2  (bases 1 to 8409)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "eLife13:RP98254"
COMMENT     Sequence Label: pAAV-CMV-HBA-RIBEYE-eGFP
FEATURES             Location/Qualifiers
     source          1..8409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(378..389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3012..3021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3018..3020,3021..3023,3024..3737)
                     /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(3018..3020,3021..3023,3024..3734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3063..3084)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3324..3343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3671..3692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3759..4347
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3812..3832)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4230..4241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4380..4397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4386..4593
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4602..4731
                     /label=AAV2 ITR
     rep_origin      4838..5293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4925..4944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5135..5156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5360..5379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5479..5501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5539..5557)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5625..5729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5730..5798,5799..6590)
                     /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(5948..5967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6761..7349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7250..7269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7672..7975
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccccacctct gctgtattgc tttgctctgc caatgccggt tcccagcagg cacatcaaca
       61 ttggccgttc tcagagctgg gatgccgctg gctggtatga gggcccttgg gagaatgctg
      121 gccctccagg gaggagaagt tctctgactt atggccctgg agaaggggtc tggtgtgagc
      181 tggtaaacca ccgtgcccag gacactgagt cctgcctgtc ccgggaagct ttttataact
      241 cactggcatc aagaaagggc tccgtgcctg acttcacctt ctacgacagc aggcaggcag
      301 tgatgtctgg ccgtggctcc gtgctgccac aagactacta tggcgaccca tccaggggca
      361 ccagggtacc caaggagcct cccttctatc gggacccagg aaccagccgg ccggtgccca
      421 gctacggagt gctgggcagc agaataccat gggagcaggt acaaggccag ttacctgctc
      481 tgcaggatgc tggccacctg taccgggaat ctgggagtaa aactgtcctg catgggcagc
      541 ggacacactg cagggctcca tctcctggga ggtatggaag ggaacaacct gattctagat
      601 tagggataga ggtacccacc tattccccca actccagcca ggtctacaac gatatctgcg
      661 aaagacccgt tgactctacc catgccagac aggtggcccc aacatgcctg gttgtggacc
      721 ccagctcgac cgcccctact gagaacagca cgggggtggc cccaggttcc ctgaatagag
      781 gctacgggcc tacccgagaa agcatccatt caaagctggc ttatgagaat tatgaagccg
      841 acttgtcaac attccaagga cccgggggca agaggaccgt ctaccctgag tttttggccc
      901 tgctgagggc agagggtgtg gcagaggcta ccctggcagc cctgttacaa caaggctttg
      961 actccccagc tgtcttggcc accctagagg atgctgacat caagtctgtt gcccccaacc
     1021 tgggccaggc ccgtgtcctg agccgcctgg tcagcagctg caggacagag atgcagtttc
     1081 ggcgacagga ccggacaggc cctccacccc gccatcggtc cagtagcttc agccaccgaa
     1141 gtgagctgct gcccaatgac acggcctcgc tggggaccac agcactgcag tttcacccag
     1201 ctggaccttt gcagacaccc tctccccgag gtggagatct aggtcgccgc ccctccagtg
     1261 caccttccca gcacctcctg gagacagcag ctacttactc tgccccagtg gtggggtccc
     1321 agactcccca cttaccctcc aactctggat atagctctcc taccccttgt gccctaacag
     1381 cacgactggc ctcctcctac ccttcacaag ctggggtagc cttgactgct aacccaggcc
     1441 cctctgtccc ccttcactca agtcccagaa cagcctattc cacctcctac acggtgccca
     1501 tggaactgct taagagggaa cgcagtgtga ccgcctctcc gctgcccagc ccccatgcca
     1561 gcccccagct cctgcggaaa cctggagcag cccccgtgga gccggctgcc ctgccaccag
     1621 tcaggcaaag tctccacaca cctcatccgc cataccagaa ggtggcccgg cggacagggg
     1681 ctcccatcat cgtgtcaacc atgctaacac cagaaccaag tatccgcccc cagatcatga
     1741 acggccccct gcacccccgc cccctagtgg cactgctgga tggcagagac tgcactgtgg
     1801 aaatgcccat cctgaaggac ctggccactg tggccttctg tgatgcacag tccactcagg
     1861 aaatccatga gaaggtattg aatgaggctg tgggcgccat gatgtaccac accatcactc
     1921 tcacccggga agacctggag aagttcaagg ccctgagagt gattgttcga atcggtagtg
     1981 gctacgacaa tgtggacatc aaggctgctg gcgaactcgg aatcgccgtg tgcaacatcc
     2041 catctgctgc agtggaggag acagcagatt cgacagtctg ccatatcctc aatctgtatc
     2101 ggcggaacac atggctgtac caggccctac gagaaggcac gcgggtacaa agcgtggaac
     2161 agatccgtga ggtcgcctca ggagctgctc ggatccgagg ggaaacactg ggcctcattg
     2221 gctttggtcg cactgggcag gcagttgctg ttcgagccaa ggcctttgga ttcagcgtca
     2281 tattttatga cccctactta caggatggga tagagcggtc tctgggcgtg caaagggtct
     2341 acaccctgca ggacttgtta tatcagagcg actgtgtctc cttgcactgc aatctgaatg
     2401 aacacaacca ccacctcatc aatgacttca ccatcaagca gatgaggcaa ggagcattcc
     2461 ttgtgaatgc agcccgaggt ggcctggtgg atgagaaagc cttagctcag gccctcaaag
     2521 agggcaggat acgaggcgct gctctggatg tgcatgagtc tgagcccttc agctttgctc
     2581 agggcccatt gaaggatgct ccaaatctca tctgcacacc acacacagcc tggtacagcg
     2641 aacaagcatc actagagatg agggaagcag ctgccacgga gatccgccga gcaatcacag
     2701 gtcgcatccc agaaagctta cgaaactgtg tcaacaaaga attctttgtt acttcaactc
     2761 cttggtcagt cattgaccag caagcaattc atcctgagct caatggtgcc acatacaggt
     2821 atccaccagg cattgtaggt gtggctccag gagggcttcc tccagctatg gaagggatca
     2881 tccctggagg catccctgtg actcacaacc tccccactgt ggcacaccct tcccaagctc
     2941 cctccccaaa ccagcccaca aaacacgggg acaatcgaga gcaccccaac gagcaaatgg
     3001 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3061 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3121 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3181 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3241 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3301 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3361 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3421 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3481 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3541 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3601 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3661 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3721 acgagctgta caagtaaagc ggccgcaagc ttatcgataa tcaacctctg gattacaaaa
     3781 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3841 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3901 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3961 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     4021 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     4081 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4141 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     4201 tgcgcgggac gcccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4261 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4321 ggatctccct ttgggccgcc tccccgcatc gataccgtcg actagagctc gctgatcagc
     4381 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4441 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4501 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4561 ggattgggaa gacaatagca ggcatgctgg ggagagatct gaggaacccc tagtgatgga
     4621 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4681 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggagt
     4741 ggcccccccc cccccccccc ctgcagcctg gcgtaatagc gaagaggccc gcaccgatcg
     4801 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5041 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5161 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5281 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5341 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5401 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5461 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5521 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5581 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5641 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5701 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5761 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5821 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5881 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5941 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6001 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6061 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6121 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6181 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6241 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6301 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6361 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6421 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6481 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6541 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6601 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6661 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6721 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6781 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6841 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6901 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6961 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7021 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7081 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7141 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7201 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7261 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7321 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7381 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7441 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7501 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7561 cgcgttggcc gattcattaa tgcaggctgc agagagggag tggccaactc catcactagg
     7621 ggttcctcag atctaggatc acgcgttcta gatcccatat atggagttcc gcgttacata
     7681 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7741 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7801 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7861 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7921 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     7981 gcggttttgg cggccgggcc cagcacccca aggcggccaa cgccaaaact ctcccccctc
     8041 ctcttcctca atctcgctct cgctcttttt ttttttcgca aaaggagggg agagggggta
     8101 aaaaaatgct gcactgtgcg gcgaagccgg tgagtgagcg gcgcggggcc aatcagcgtg
     8161 cgccgttccg aaagttgcct tttatggctc gagcggccgg ccgcggcggc gccctataaa
     8221 acccagcggc gcgacgcgcc accaccgccg agaccgcgtc cgccccgcga gcacagagcc
     8281 tcgcctttgc cgatccgccg cccgtccaca cccgccgcca ggtaagcccg gccagccgac
     8341 cggggcaggc ggctcacggc ccggccgcag gcggccgccc aaagaattcg gcacgaggaa
     8401 aggcctcct
//
