LOCUS       sequence-491769-        6144 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for GRIN2A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  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  JOURNAL   Unpublished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 6144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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-5'-GRIN2A
FEATURES             Location/Qualifiers
     source          1..6144
                     /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"
     regulatory      707..716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3188..3197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      3622..4077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3709..3728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3919..3940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4094..4113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4213..4235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4273..4291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4359..4463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4464..4532,4533..5324)
                     /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(4682..4701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5495..6083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5984..6003
                     /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 ccatgggcag
      721 agtgggctat tggaccctgc tggtgctgcc ggcccttctg gtctggcgcg gtccggcgcc
      781 gagcgcggcg gcggagaagg gtccccccgc gctaaatatt gcggtgatgc tgggtcacag
      841 ccacgacgtg acagagcgcg aacttcgaac actgtggggc cccgagcagg cggcggggct
      901 gcccctggac gtgaacgtgg tagctctgct gatgaaccgc accgacccca agagcctcat
      961 cacgcacgtg tgcgacctca tgtccggggc acgcatccac ggcctcgtgt ttggggacga
     1021 cacggaccag gaggccgtag cccagatgct ggattttatc tcctcccaca ccttcgtccc
     1081 catcttgggc attcatgggg gcgcatctat gatcatggct gacaaggatc cgacgtctac
     1141 cttcttccag tttggagcgt ccatccagca gcaagccacg gtcatgctga agatcatgca
     1201 ggattatgac tggcatgtct tctccctggt gaccactatc ttccctggct acagggaatt
     1261 catcagcttc gtcaagacca cagtggacaa cagctttgtg ggctgggaca tgcagaatgt
     1321 gatcacactg gacacttcct ttgaggatgc aaagacacaa gtccagctga agaagatcca
     1381 ctcttctgtc atcttgctct actgttccaa agacgaggct gttctcattc tgagtgaggc
     1441 ccgctccctt ggcctcaccg ggtatgattt cttctggatt gtccccagct tggtctctgg
     1501 gaacacggag ctcatcccaa aagagtttcc atcgggactc atttctgtct cctacgatga
     1561 ctgggactac agcctggagg cgagagtgag ggacggcatt ggcatcctaa ccaccgctgc
     1621 atcttctatg ctggagaagt tctcctacat ccccgaggcc aaggccagct gctacgggca
     1681 gatggagagg ccagaggtcc cgatgcacac cttgcaccca tttatggtca atgttacatg
     1741 ggatggcaaa gacttatcct tcactgagga aggctaccag gtgcacccca ggctggtggt
     1801 gattgtgctg aacaaagacc gggaatggga aaaggtgggc aagtgggaga accatacgct
     1861 gagcctgagg cacgccgtgt ggcccaggta caagtccttc tccgactgtg agccggatga
     1921 caaccatctc agcatcgtca ccctggagga ggccccattc gtcatcgtgg aagacataga
     1981 ccccctgacc gagacgtgtg tgaggaacac cgtgccatgt cggaagttcg tcaaaatcaa
     2041 caattcaacc aatgagggga tgaatgtgaa gaaatgctgc aaggggttct gcattgatat
     2101 tctgaagaag ctttccagaa ctgtgaagtt tacttacgac ctctatctgg tgaccaatgg
     2161 gaagcatggc aagaaagtta acaatgtgtg gaatggaatg atcggtgaag tggtctatca
     2221 acgggcagtc atggcagttg gctcgctcac catcaatgag gaacgttctg aagtggtgga
     2281 cttctctgtg ccctttgtgg aaacgggaat cagtgtcatg gtttcaagaa gtaatggcac
     2341 cgtctcacct tctgcttttc tagaaccatt cagcgcctct gtctgggtga tgatgtttgt
     2401 gatgctgctc attgtttctg ccatagctgt ttttgtcttt gaatacttca gccctgttgg
     2461 atacaacaga aacttagcca aagggaaagc accccatggg ccttctttta caattggaaa
     2521 agctatatgg cttctttggg gcctggtgtt caataactcc gtgcctgtcc agaatcctaa
     2581 agggaccacc agcaagatca tggtatctgt atgggccttc ttcgctgtca tattcctggc
     2641 tagctacaca gccaatctgg ctgccttcat gatccaagag gaatttgtgg accaagtgac
     2701 cggcctcagt gacaaaaagt ttcagagacc tcatgactat tccccacctt ttcgatttgg
     2761 gacagtgcct aatggaagca cggagagaaa cattcggaat aactatccct acatgcatca
     2821 gtacatgacc aaatttaatc agaaaggagt agaggacgcc ttggtcagcc tgaaaacggg
     2881 gaagctggac gctttcatct acgatgccgc agtcttgaat tacaaggctg ggagggatga
     2941 aggctgcaag ctggtgacca tcgggagtgg gtacatcttt gccaccaccg gttatggaat
     3001 tgcccttcag aaaggctctc cttggaagag gcagatcgac ctggccttgc ttcagtttgt
     3061 gggtgatggt gagatggagg agctggagac cctgtggctc actgggatct gccacaacga
     3121 gaagaacgag gtgatgagca gccagctgga cattgacaac atggcgggcg tattctacat
     3181 gctggctgcc gccatggccc ttagcctcat caccttcatc tgggagcacc tcttctactg
     3241 gaagctgcgc ttctgtttca cgggcgtgtg ctccgaccgg cctgggttgc tcttctccat
     3301 cagcaggtaa gtatcaaggt tacaagacag gtttaaggag accaataagc tattattcgt
     3361 ataggatact ttatacgaag ttattaagtt atggagaccg gtgtggcacg tgcggaccga
     3421 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3481 cactgaggcc gggcgaccaa aggtcgcccg gcctcagtga gcgagcgagc gcgcagctgc
     3541 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3601 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3661 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     3721 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3781 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     3841 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3901 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     3961 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4021 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4081 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4141 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4201 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4261 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4321 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4381 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4441 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4501 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4561 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4621 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4681 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4741 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4801 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4861 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4921 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4981 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5041 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5101 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5161 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5221 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5281 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5341 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5401 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5461 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5521 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5581 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5641 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5701 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5761 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5821 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5881 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5941 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6001 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6061 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6121 tttgctggcc ttttgctcac atgt
//
