LOCUS       sequence-489073-        6619 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for GRIN2B expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6619)
  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  JOURNAL   Unpublished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 6619)
  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            "Unpublished"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-5'-GRIN2B
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2649..2917
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2930..3141)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   3198..3327
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4148..4939,4940..5008))
                     /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     4771..4790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5009..5113)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5181..5199
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5237..5259)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5359..5378
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5395..5850)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5532..5553)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5744..5763
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5925..6065
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5936..6065
                     /label=AAV2 ITR
ORIGIN
        1 atgcagctgt agatacctct gctgatggag aagaccatgc caggcttgcc agaacagaca
       61 cccataaagc aatgtcggaa ctgccaatag aaaaggtgtt cgcagatgaa ggtgatgagg
      121 ctgagagcca tggccgcccc caacatgtag aagacccctg ccatgttgtc aatgtccagc
      181 tggctgctca tgacctcatt cttctcattg tgacaaatgc cagtgagcca gagagcttcc
      241 agttcttcca tctccccatc tccaaagagc tgcaggatag caaggtccac ctggcgcttc
      301 cacccagaat ctttttggat ggcaatgcca tagccagtgg aagcaaagac cttcccactg
      361 ccaatggtca ccagcttgca gccttcatct ctgcctgcca tatagttcag cactgctgca
      421 tcatagatga aggcatccag tttccctgtt ttcagggaga gcaatgcatc atctacaccc
      481 ctctggttga actttcccat gtaggcatgc atttctgcat agttattgcg aatatttctc
      541 tctgtgctgc cgttgggcac ggtcccaaag cggaaagggg gtgagaagtc attaggtctc
      601 tggaactttt tgtcgctcag gccagaaacc tggtccacat attcctcttg gatcatgaag
      661 gcagctaagt tggcagtgta gctggccagg aagatgacag caaagaaggc ccacactgac
      721 accatgatct tggaggtggt cccctttggg ttctgcacag gtacggagtt gttaaacacc
      781 agaccccaga gcaaccaaat agctttgccg atggtgaaag agggtccacc aggctctctg
      841 ccatcagcga ggcacctgtt ataacccaca gggctgaagt actcaaagac aaagacagcc
      901 acggctgaga cgatgagcag catcacaaac atcatcaccc atacgtcagc gctgaatggc
      961 tctaagaagg cagaaggtga gacagtccca ttgctgcgtg acaccatgac actgatgcct
     1021 gtctctatga agggcacaga gaagtcgacc acctccgatc gttcctcatt gatggtgagt
     1081 gagcccactg ccatgtaggc cctcttcatg accacctctc caatcatacc attccaggtt
     1141 ccattgattt tcttcccatg cttgccattg gtaaccaggt aaaggtcata ggtgaacttc
     1201 acagatttag aaattttctt aaggatgtca atacagaacc ccttgcagca ttttttgatg
     1261 taacccggct cctcgtctgt tttattctca gtgactatgc gtttttggca ggggactgtg
     1321 ttcctcatgc aggttccact cagagggtcc acactttcca caatgacaaa tggtgcctcc
     1381 tccagggtca caatgctcag atggtcatcc tcctgctctt cagtctctgg acacattcgg
     1441 ggccacacat agtacttcat ctgcagggac ttgtctttcc acttccccac cctttcccac
     1501 ttcctctcct tgttcagaag aattatcacc agtttcgggt gcatctggta gccatcttca
     1561 ctgaaggaca aattcctccc ctcaaaagtg acattgatca gatacctatt tagcatattg
     1621 gactggtaga ttctcttctc gtgggtgttg taacaactgc ttttgggctc agggatgaag
     1681 ctgtgctcag acagcatgtc agaagcagca gtggtgatta tggcaattcc atctctcact
     1741 ctggcgggga ggccatagtc ccattcatca tatgatacag agatgagccc agtggggaac
     1801 tccgcaggca ctgtgtctgt atcccctgcc accagactgg gcacgatcca cgtgtagcca
     1861 tagccagtca gccctactga gttggccact tcaaagatgt aggtggcttc ttccttggta
     1921 cagtaaagaa gaatgatggg gctttgaagt ttcttgagct gattctggat cttagaatct
     1981 ccatcgtcca gggacatgtc cagtaggagg acctcctcta gctcccagcc cacaaagcta
     2041 ttctcaatgg tgctgcggat cttgtttaca aagtcctggt agccagggaa ataggtggtg
     2101 acgatagaaa agatgtacca gtcatattct tccatgatgt tgagcattac ggaagcttgc
     2161 tgttcaattg atgggccaaa ctggaagaac atggaggatt catccttatc tgccattatc
     2221 atagaggagc ccccgtggat gcccaggatg ggggtgagag tctgtgctga aatgaaatcg
     2281 aggatctggg cgatggcttc ctggtctgtg tcatcagcaa acaccacccc ctggatcttc
     2341 cggtcagaca tgagatcaca gatgcgggtg atgatgctct ttgggtcggt ctcattcatg
     2401 gctaccagtt ccacccgggg taccacggag agatggtgga aatcatcttt ctcgtgggca
     2461 tccttgatgg ccacctcgtc ggaagtgccc acgaggatga cagcaatgcc aatgctgggg
     2521 gggctcttct gagaacgagc tctgctgcct gacacggcca ggacggccaa caccaaccag
     2581 aacttgggag aacagcactc cgctctgggc ttcatggtgg ccgtacggta ggcgccggtc
     2641 acagcttgga tctgtaacgg cgcagaacag aaaacgaaac aaagacgtag agttgagcaa
     2701 gcagggtcag gcaaagcgtg gagagccggc tgagtctagg taggctccaa gggagcgccg
     2761 gacaaaggcc cggtctcgac ctgagcttta aacttaccta gacggcggac gcagttcagg
     2821 aggcaccaca ggcgggaggc ggcagaacgc gactcaaccg gcgtggatgg cggcctcagg
     2881 tagggcggcg ggcgcgtgaa ggagagatgc gagcccctcg aagcttcagc tgtgttctgg
     2941 cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc
     3001 ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc
     3061 gccaccttct ctaggcaccc gttcaattgc cgacccctcc ccccaacttc tcggggactg
     3121 tgggcgatgt gcgctctgcc cactgacggg caccggagcg atcgcagatc ctctagagtc
     3181 gacacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3241 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     3301 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5161 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     5221 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5281 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5341 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     5401 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5461 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     5521 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5581 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     5641 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5701 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5761 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5821 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     5881 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     5941 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     6001 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     6061 ttcctgcggc cgctcggtcc gcacgtgcca caccggtctc cataacttaa taacttcgta
     6121 taaagtatcc tatacgaata atagcttatt ggtctcctta aacctgtctt gtaaccttga
     6181 tacttacctg gactgggtgg tgaagggtgg gttgtcacag tcgtagagcc catcgatgga
     6241 gctggcactg ccaatactat ggggccggtg gtgatggtgg tagtgatctt ggtacacgtt
     6301 gctgtccttc agctgcaggt tcccgaacgt tctctctacc tcactgatgt agtcactgaa
     6361 gaggttctcc tcacagggcg ggttgttgta ggatttgcag tcagaatgcg tgaagctgcg
     6421 gcggtgctct gagatgtcat agacggatga ctcccgtcgg atgaagtcca gggcgctctg
     6481 cggtgagcca ttcacaccag acaggttagc catgttcttg gccgtgcgca gcaggcgcag
     6541 gatgttggag tgtgtgttgt tcatggttgc ggtgggggag ttcattacag actggcgctc
     6601 ctcgatcgcc accccatgg
//
