LOCUS       sequence-489075-        5267 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for SYNGAP1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5267)
  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  JOURNAL   Unpublished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 5267)
  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            "Unpublished"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'-SYNGAP1
FEATURES             Location/Qualifiers
     source          1..5267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(228..439)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   496..625
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(687..1275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(767..786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1446..2237,2238..2306))
                     /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     2069..2088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2307..2411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2479..2497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2535..2557)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2657..2676
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2693..3148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2830..2851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3042..3061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3223..3363
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3234..3363
                     /label=AAV2 ITR
     CDS             complement(5151..5177)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             join(5214..5267,1..215)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
ORIGIN
        1 agggtcaggc aaagcgtgga gagccggctg agtctaggta ggctccaagg gagcgccgga
       61 caaaggcccg gtctcgacct gagctttaaa cttacctaga cggcggacgc agttcaggag
      121 gcaccacagg cgggaggcgg cagaacgcga ctcaaccggc gtggatggcg gcctcaggta
      181 gggcggcggg cgcgtgaagg agagatgcga gcccctcgaa gcttcagctg tgttctggcg
      241 gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc
      301 ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc
      361 caccttctct aggcacccgt tcaattgccg acccctcccc ccaacttctc ggggactgtg
      421 ggcgatgtgc gctctgccca ctgacgggca ccggagcgat cgcagatcct ctagagtcga
      481 cacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
      541 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc
      601 gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac
      661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
      901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     1921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2401 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     2461 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     2521 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2581 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2641 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta
     2701 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2761 caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg
     2821 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2881 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt
     2941 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3001 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3061 gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3121 gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa
     3181 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg
     3241 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     3301 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     3361 cctgcggccg ctcggtccgc acgtgccaca ccggtctcca taacttaata acttcgtata
     3421 aagtatccta tacgaataat agcttattgg tctccttaaa cctgtcttgt aaccttgata
     3481 cttaccttgc acagggccaa ctcacagcac attcgcaggt tggcctggtg ctctgccaaa
     3541 ctggatgctg tgcacttgat agggtctacc tcgcagtttt cctcagattc atacagagca
     3601 cggatgaatt ctccaatggc atccttgagg tatttctgac caatcagtct catatactct
     3661 tctatggctt tagtggcaag cgtgttctcg cggaatatga ggtgctcccg ttccatgaac
     3721 cggtctacct cagacatggc catgtctgaa aggaagtcct tggccttgcc tgtactctgc
     3781 aggatgtgaa ctagtgcact ggcaacctcc tccttgcctt tgacattcag ggcgggctcc
     3841 aagactgcac acagcatccg ataatggttg gtgacatact ctgcaaactc tttatatagc
     3901 tccatgggca agatgctcat tgtctggtaa cgtgctttca gccgcacagc cgggcaacct
     3961 cctttgccct tgccccctga gccacccccc gagccccctc cccctcccga acccatgccc
     4021 ccagatcccc cactgcctgt tggcagggtt acagggtacc actgctctgt gaagtggcgc
     4081 ccagccaggg tggccactgg cacagtcacc aggccgacat agcctgcctt gtccttcttg
     4141 cgctttttgt ctgagtcacg gtacagatgc agccgcaggg cacggacagc cggcaggttg
     4201 ttaaactcga agtgctcgcc ccagaagacg gtgtccccag aggcagagcg gggcttggag
     4261 gtggtgcgtg catacagcat gtcatccagg cagagctcac agtagtaccg cttcttgggg
     4321 ggcagctccc gggcctctat gatccacagc tttagcacat tgtctacccg gcggctgttg
     4381 tccttgttgg gctttactgc ccgctgcaga ttctcaatcc atttgtctct ttcggccgca
     4441 gaccgacagg caaagcattt tgttcctgat gaagttgtta cctcaaaaca gaactcctgg
     4501 cccaggatgg agctgtgcac tggcttgata atggaatctt catccaagtt gagctccaat
     4561 gcctcagctg cactgctagg actcagcaag gactcatgag agtgtgactc cttaaagctt
     4621 tgcatcagcc gggcccggtc atggtcagca cttcggaagc gaggcaggat ctggcgaaag
     4681 ctgctggtcc ggtcaagttt gggttgtgac ttcgttcgtt tgatggagct ttttagccgt
     4741 cggctcagga agccttgcga gggccggaag ggcgcagcag gggcaccctc catgctgtac
     4801 tgcttccccc ctgggacact cttcctcctc gagcgaccca agtggtattc atgctcgccg
     4861 tggggccgcc cctccaccgg cacggagact gtgcgtctca gcagtttgtt gcgactggac
     4921 tcgctcctcc ggtcccggat cagtaggggg tgtatctcat cctcatccag catgagcagc
     4981 tggttccccg agatgatgca gaaccgaggg ttccaaccag gacgatcata cggggaatga
     5041 acgtattggg ttcggtgcat agagggtccc cgtacatctc tgaagggggc ataggacatc
     5101 gcggggatgc tcccccgatg gatggaggct cgagacctgc tcatagatcc agcatagtca
     5161 ggtacgtcat aagggtacat ggtggccgta cggtaggcgc cggtcacagc ttggatctgt
     5221 aacggcgcag aacagaaaac gaaacaaaga cgtagagttg agcaagc
//
