LOCUS       sequence-489081-        6294 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for CLASP1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  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  JOURNAL   Unpublished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 6294)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 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            "Unpublished"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'-CLASP1
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(269..295)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             332..600
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(613..824)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   881..1010
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1072..1660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1152..1171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1831..2622,2623..2691))
                     /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     2454..2473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2692..2796)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2864..2882
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2920..2942)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3042..3061
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3078..3533)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3215..3236)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3427..3446
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3608..3748
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3619..3748
                     /label=AAV2 ITR
ORIGIN
        1 gcgccttgaa ccgatcctgc agccgggtca ccagggcgga caggatgtcc atgcccagca
       61 gaaccacctt gtaattgcta gagttcaccc aagaggtagc aagtccatcc acaagtttat
      121 ctaacatggt ctggtcatgc tcaaggtcag cagacttctg tttgtctgag aaatagtcta
      181 tcagttcttg gccaacctgc aatcgtttcc ccacatcctt ctgcaacacc tgcgccaggc
      241 aggactccat gcgaggctcc atagatccag catagtcagg tacgtcataa gggtacatgg
      301 tggccgtacg gtaggcgccg gtcacagctt ggatctgtaa cggcgcagaa cagaaaacga
      361 aacaaagacg tagagttgag caagcagggt caggcaaagc gtggagagcc ggctgagtct
      421 aggtaggctc caagggagcg ccggacaaag gcccggtctc gacctgagct ttaaacttac
      481 ctagacggcg gacgcagttc aggaggcacc acaggcggga ggcggcagaa cgcgactcaa
      541 ccggcgtgga tggcggcctc aggtagggcg gcgggcgcgt gaaggagaga tgcgagcccc
      601 tcgaagcttc agctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
      661 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
      721 atcactttcc cagtttaccc cgcgccacct tctctaggca cccgttcaat tgccgacccc
      781 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga
      841 gcgatcgcag atcctctaga gtcgacacgc gtgcggccgc aggaacccct agtgatggag
      901 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
      961 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc
     1021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2821 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     2881 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2941 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3001 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     3061 actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     3121 ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc
     3181 aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt
     3241 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3301 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3361 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3421 aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg tagcggtcac
     3481 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg
     3541 ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     3601 aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     3661 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     3721 gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg ccacaccggt
     3781 ctccataact taataacttc gtataaagta tcctatacga ataatagctt attggtctcc
     3841 ttaaacctgt cttgtaacct tgatacttac cttctttcct ttctgaccag tttgaactag
     3901 cacagtggtt gagaacttct gctacatcct cagtctgccg cagataatgg ggaatgccac
     3961 cattcctgga gccatatgag cgctcagagc aaacacttga ggcatcactg ttggcatcat
     4021 cgtcagaata catcccatac ggctcatatc tcctcctcac aggcttcttc aaagcatcag
     4081 caacagcagc ttcaagatct gtacttgtgc tcagaactct catggcattc acagaaccag
     4141 gtattcttcc tggctggcca agcccaaacc gatctgactg cccaacagat tcagcagtgg
     4201 agagagcact agtggacctg gaatgacgtc gagaggcaag tggaggaaag ccccgagcag
     4261 ggcttgtatc tcggctgctc tcacggctgg tatcgcggct gcacccctga ctcatgctgg
     4321 gtcgagggat acggctgctc cgtgctaatc ctattcggtt tggacttgtt tcccggctac
     4381 atccctggct cctgggaatc ttgcttcgct tttctgaaga cggtgtcaca ggtgggcctc
     4441 gtgaggagcc cccagtaagt ccaccataac cacttcccaa caattttcct ggggaacttg
     4501 accggctgcc agcaccagca ggattagcag atctggatcg ctgggactgt gaaaccactt
     4561 tagcgcgact gcggccccgg ttatcaggtg tagaggcgac gttggtggca ctcccagagc
     4621 tttgccgtct tgtgcggatc cgacctaagg atgcgtatga ccctggaggc aaagctgctg
     4681 cagagctgaa aggcgtcgtg cccgaagatg aggagacttt ggatttggca ctggctgctg
     4741 cgttcacatc aatatcactt cgagatcgct ggagggaccc agtcgttgac acagatttgg
     4801 tactaactgt agaagctcta gatgtggtac ttccagtagg acttcttttg gcagacagcg
     4861 gacgatttag actctcttga gagctggaag atgagcggtc tgactgaggc agagacacta
     4921 tgctgtctga gttcttcagg tgggactgca gggctttctg gtaggaggac tccaaggtgt
     4981 ggtacaagtg ctctgcttct ctgctgaagt gactgtggaa accccagtaa cattttctgg
     5041 cttctattct tgcttcggaa tcagcatcat gtattccctt ctttattgtt tcagctaata
     5101 ctgatatgtg tcgttctagt gaatgtgtct gccattcttg taaaagcaaa tctaaaaatt
     5161 caaaacagcg ccttctaact gcgacagact tagaggtaca gttgcttgtt atgacaggta
     5221 ttaacctagg gatgtgtgtg tgccgaataa ttaacctaac agctacaaca ccagatgtgg
     5281 ccataatttt ggcactgttt ggaattaaat taaagatagt tggcataatg gcttcagctc
     5341 catggtcaaa cttattcccc agaactgatg acagatgccc caacgtgata caagcctccc
     5401 gcactacctg agaccgcagg tccttagcag agagtttaaa ggctccatcc aaaagacgca
     5461 aatgttgaaa gaagttatca tactcagcag caccagccaa aagtaaagat ctaatctttt
     5521 ttagagcatt tactctctgc tcccaatcat gcttgtcatc agataatatt tccctaattt
     5581 tgtttatgga ttcctcaagg tctcggctgg aataaatctg tactacaggt acatcatcaa
     5641 atgctttaat aaaatcctct tcatcaacag caccagctcc ttcttttgca gctgaagact
     5701 tggatccaag ggtggatgaa ccaagccggc gggtggttcc cattccaacg tttctccgag
     5761 aacttggtgg agccttggat gatgtagaac tagcagagga aggtctgtta ccatccacag
     5821 aatcttcatc gtcgaaattt ttatcatttg cagattgtat catgtttcca gatttctgga
     5881 cttcatcaaa ttttgtaaaa attacattca accgggactg tggcaatcct tttttactga
     5941 gatctgccct cacacgttct cctacatgtc tgtaaatttc cactaagctg tttattgctg
     6001 catctcgaac ctggctgttt ggatctccaa gtaagttgca tatatgtggc acaatcttgc
     6061 ttagtgttaa agtctgtgct ccagaggcat tgagtgttgc tataaggcag agacagatgc
     6121 cttctcgagt acggaaattc ttgtgtttga agcctccaag cattctgtcc catacgtact
     6181 ggggattagc agcttgatcc atgatcttta gcagcagagt ttggtcctgc tccctcacag
     6241 agtctttagc atctcctagt ctgtctatta gacttggcag cactgtgccg atct
//
