LOCUS       sequence-491930-        6590 bp    DNA     circular SYN 05-NOV-2025
DEFINITION  5' AAVLINK plasmid for UNC13A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6590)
  AUTHORS   Zhonghua Lu Lab
  TITLE     AAVLINK: A Potent DNA-Recombination Method for Large Cargo Delivery 
            in Gene Therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6590)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EF1alpha-HA-5'-UNC13A
FEATURES             Location/Qualifiers
     source          1..6590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   3853..3993
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3853..3982
                     /label=AAV2 ITR
     rep_origin      4068..4523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4155..4174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4365..4386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4540..4559)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4659..4681
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4719..4737)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4805..4909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4910..4978,4979..5770)
                     /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(5128..5147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5941..6529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6430..6449
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctat gtctctgctt tgcgttggag tcaaaaaagc
      781 caagtttgat ggtgcccaag agaaattcaa cacgtacgtg accctgaaag tgcagaatgt
      841 caagagcacg accatcgcgg tgcggggcag ccagcccagc tgggagcagg atttcatgtt
      901 cgagattaac cgtctggatt tgggactgac ggtggaggtg tggaataagg gtctcatctg
      961 ggacacaatg gtgggcactg tgtggatccc actgaggacc atccgccagt ccaatgagga
     1021 gggccctgga gagtggctga cgctggactc ccaggtcatc atggcagaca gtgagatctg
     1081 tggcaccaag gaccccacct tccaccgcat cctcctggac acgcgctttg agctaccctt
     1141 agacattcct gaagaggagg ctcgctactg ggccaagaag ctggagcagc tcaatgctat
     1201 gcgggaccag gatgaatatt ccttccaaga tgagcaagac aagcctctgc ctgtccccag
     1261 caaccagtgc tgcaactgga attattttgg ctggggtgag cagcacaacg atgaccccga
     1321 cagtgcagtg gatgatcgtg acagtgacta ccgcagtgaa acgagcaaca gcatcccgcc
     1381 gccctattat actacgtcac aacccaacgc ctcagtccac caatattctg ttcgcccacc
     1441 acccctgggc tcccgggagt cctacagtga ctccatgcac agttacgagg agttctctga
     1501 gccacaagcc ctcagcccca cgggtagcag ccgctatgcc tcttccgggg agctgagcca
     1561 gggaagctct cagctgagcg aggacttcga ccctgacgag cacagcctgc agggctccga
     1621 catggaggat gagcgggacc gggactccta ccactcctgc cacagctcgg tcagctacca
     1681 caaagactcg cctcgctggg accaggatga ggaagagctg gaggaggacc tggaggactt
     1741 cctggaggag gaggagctgc ctgaagatga ggaggagctg gaggaggagg aggaggaggt
     1801 gcctgacgat ttgggcagct atgcccagcg tgaagacgta gctgtggctg agcccaaaga
     1861 cttcaaacgc atcagcctcc cgccagctgc cccagggaag gaggacaagg ccccagtggc
     1921 acccaccgag gcccccgaca tggccaaggt ggcccccaag ccagccacgc ccgacaaggt
     1981 gcctgcagct gagcagatcc ctgaggctga gccacccaag gacgaggaga gtttcaggcc
     2041 gagagaggat gaggaaggcc aggaggggca ggactccatg tccagggcca aggccaactg
     2101 gctgcgtgcc ttcaacaagg tgcggatgca gctgcaggag gcccggggag aaggagagat
     2161 gtctaaatcc ctatggttca aaggcggccc agggggcggt ctcatcatca tcgacagcat
     2221 gccagacatc cgcaagagga aacctatccc actcgtgagc gacttggcca tgtccctggt
     2281 ccagtccagg aaagcgggca tcacctcggc cttggcctcc agcacgttga acaacgagga
     2341 gctgaaaaac cacgtttaca agaagaccct gcaagcctta atctacccca tctcgtgcac
     2401 gacgccacac aacttcgaag tgtggacggc caccacgccc acctactgct acgagtgcga
     2461 ggggctgctg tggggcatcg cgaggcaggg catgcgctgc accgagtgcg gtgtcaagtg
     2521 ccacgagaag tgccaggacc tgctcaacgc cgactgcctg cagcgggctg cggagaagag
     2581 ctccaagcac ggggcggagg accggacaca gaacatcatc atggtgctca aggaccgcat
     2641 gaagatccgg gagcgcaaca agcccgagat cttcgagctc atccaggaga tcttcgcggt
     2701 gaccaagacg gcgcacacgc agcagatgaa ggcggtcaag cagagcgtgc tggacggcac
     2761 gtccaagtgg tccgccaaga tcagcatcac cgtggtctgc gcccagggct tgcaggcaaa
     2821 ggacaagaca ggatccagtg acccctatgt caccgtccag gtcgggaaga ccaagaaacg
     2881 gacaaaaacc atctatggga acctcaaccc ggtgtgggag gagaatttcc actttgaatg
     2941 tcacaattcc tccgaccgca tcaaggtgcg cgtctgggac gaggatgacg acatcaaatc
     3001 ccgcgtgaaa cagaggttca agagggaatc tgacgatttc ctggggcaga cgatcattga
     3061 ggtgcggacg ctcagcggcg agatggacgt gtggtacaac ctggacaagc gaactgacaa
     3121 atctgccgtg tcgggtgcca tccggctcca catcagtgtg gagatcaaag gcgaggagaa
     3181 ggtggccccg taccatgtcc agtacacctg tctgcatgag aacctgttcc acttcgtgac
     3241 cgacgtgcag aacaatgggg tcgtgaagat cccagatgcc aagggtgacg atgcctggaa
     3301 ggtttactac gatgagacag cccaggagat tgtggacgag tttgccatgc gctacggcgt
     3361 cgagtccatc taccaagcca tgacccactt tgcctgcctc tcctccaagt atatgtgccc
     3421 aggggtgcct gccgtcatga gcaccctgct cgccaacatc aatgcctact acgcacacac
     3481 caccgcctcc accaacgtgt ctgcctccga ccgcttcgcc gcctccaact ttgggaaaga
     3541 gcgcttcgtg aaactcctgg accagctgca taactccctg cggattgacc tctccatgta
     3601 ccggaataac ttcccagcca gcagcccgga gagactccag gacctcaaat ccactgtgga
     3661 ccttctcacc agcatcacct tctttcggat gaaggtacaa gaactccaga gcccgccccg
     3721 agccagccag gtaagtatca aggttacaag acaggtttaa ggagaccaat aagctattat
     3781 tcgtatagga tactttatac gaagttatta agttatggag accggtgtgg cacgtgcgga
     3841 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3901 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3961 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4021 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4141 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4261 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4381 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4441 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4501 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4561 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4621 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4681 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4741 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4801 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4861 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4921 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4981 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5041 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5101 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5161 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5221 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5281 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5341 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5401 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5461 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5521 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5581 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5641 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5701 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5761 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5821 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5881 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5941 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6001 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6061 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6121 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6181 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6241 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6301 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6361 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6421 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6481 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6541 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
