LOCUS       sequence-476890-        6351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for DIP2A expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  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  JOURNAL   Unpublished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 6351)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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            "Unpublished"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' DIP2A
FEATURES             Location/Qualifiers
     source          1..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(438..464)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             501..769
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(782..993)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   1038..1167
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1229..1817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1309..1328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1988..2779,2780..2848))
                     /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     2611..2630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2849..2953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3021..3039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3077..3099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3199..3218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3235..3690)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3372..3393)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3584..3603
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3765..3905
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3776..3905
                     /label=AAV2 ITR
ORIGIN
        1 gagccctcat cttctgaggc agacgacgtg tctggagggg tgtaggtttc cacagacgaa
       61 tgcacaagga cagaacgtct cttcgaaggc ataggcatct tcctctcttt gtatttggcc
      121 aaagctgctt gcacggcttc agtgtggaca tctgaccgga agcgctcatc cctcgaggcg
      181 gtgggccgag acttctgctg cttgggtgca gcggccgtgg tttgtgaggg cccaggaatt
      241 ctattctctg cttgcagaga tgggtctatt ccttgaataa gcggtatata acgtgcaagc
      301 agctttgccc ttttcttttc atatcctttt tgagtgatgt caccttccga cagctccagc
      361 tccagctcag ccaggctctc ccgcacctcg gcaggcagcg gcgccgcctc cagcgggcac
      421 ccgcggtcag cagatccagc atagtcaggt acgtcataag ggtacatggt ggccgtacgg
      481 taggcgccgg tcacagcttg gatctgtaac ggcgcagaac agaaaacgaa acaaagacgt
      541 agagttgagc aagcagggtc aggcaaagcg tggagagccg gctgagtcta ggtaggctcc
      601 aagggagcgc cggacaaagg cccggtctcg acctgagctt taaacttacc tagacggcgg
      661 acgcagttca ggaggcacca caggcgggag gcggcagaac gcgactcaac cggcgtggat
      721 ggcggcctca ggtagggcgg cgggcgcgtg aaggagagat gcgagcccct cgaagcttca
      781 gctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta
      841 tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc
      901 agtttacccc gcgccacctt ctctaggcac ccgttcaatt gccgacccct ccccccaact
      961 tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag cgatcgcaga
     1021 tccacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     1081 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     1141 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
     1201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3001 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3061 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3121 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3181 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     3241 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3301 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     3361 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3421 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     3481 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3541 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3601 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3661 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     3721 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     3781 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     3841 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     3901 ttcctgcggc cgctcggtcc gcacgtgcca caccggtctc cataacttaa taacttcgta
     3961 taaagtatcc tatacgaata atagcttatt ggtctcctta aacctgtctt gtaaccttga
     4021 tacttacctt ccagaaagcg ctgtttggtt tcagaaatgt gaatccctcc gagaggagcc
     4081 ttgggcaagg tgttggcagg aaccagggcc agacagtaca cgcccacctg gtggatgcta
     4141 tcaatggcct gcagcacacg gctcatccac tggaagctgt cctcctccga ggcatccggc
     4201 cgctgctcag ccaccaggac aatccggtcg tcgtgcagca cggtcacaga gaacacagcg
     4261 atcctgcctc tgtagacaaa cttcatgggc tccacggcca gtgcggtggc cacaacgtca
     4321 tctgcattgt gtctgcgaac tccagtgacc atcagcccgt ccagtttgcc cacgatgaag
     4381 accaggttgt caggcccgat gaagcccagc aggcctgtcc tggtgaatgg cctgtcaaag
     4441 atgggtgctc ctcctgtggt gaccggaact gcctcaaaca cattcttcgt gattccaagc
     4501 aatccatagt acgctgtgcc agttgcactg gaactgacgc atatttctcc cacttcatca
     4561 gttttacaaa gataaggggt accttctaac ttcacaacac atacattagc tccaggcatc
     4621 acctgaccaa cgtcctgaac agtaaggact gacaactttt cttcagtatc cactctgata
     4681 acaccataac ttagaccgtt catcgatagg actgcttttc ttggaggtgg tcctcccaga
     4741 tcaggtggcc tgcggatggc gacagtcagc gcctcaggag aacttgcaca aggacagatg
     4801 acctctggcc tcagacctct ggactggaag acgttgagga aggcgtcaca ggaggagatc
     4861 gaccacgggt tggcaccatc ggccacaatc agcatgcgca gtgagctgag gctgacgtcc
     4921 ctctggcccc gctgagctag gagagaccag tgcatgtctc gcgacttcac cagcgcggcc
     4981 cgagctttat agaagcacac tttctggatc caggagagtg ggttcgcctt catcagcgcg
     5041 taggggacgc tgaccacgtg catcctgttc atgacgcttg ttaacacgcc atgccacaga
     5101 ccagcatccc ttttgaaatc cagcacgttt gttaatgttt cagcttctga gtacccgcac
     5161 gcctgggtca gagcccggca ctgtgccagc agggatgcgt gggacactgt gacccccacc
     5221 gtactgcctt ctttgctggt tttatactca atgtaggcag tcccagtccc tgtgtcctgg
     5281 gccagagggt gccagtcctt tgggggcttg gctagatgct tcccatcaat cactagccag
     5341 gagagcgggg gccaaccttt gaaagctgcc acctctcctg tctgtgcctt ggggaggcct
     5401 ttctgacaag cgtctgtggt cagggccaag aagactccac agctgcccag cagaaaccca
     5461 acctgctggc tgcctgcatc ctttcttgtt aatggcactt ctatggggac aggaaccagc
     5521 tctgccagga gacacccata aaatgcaacc atgaacatca cagggtcact attcggaaac
     5581 acgagcgcca ctctgtctcc aggtttaagt agaggttcat tcttacttgt cagtttatta
     5641 agtagagtat aagctagttt taaactccga ctccaaagtt taccataggt gagagtgtag
     5701 acggctttcc cagttgtatc caaggcagtc agacaggggg atttgggctg tgttgtgccc
     5761 cagcgctgca aggtggccaa cagcgacggc ggtcggggga cacctgcagt gagaggctcc
     5821 cctctcagca cactcgtctc gcttccctca ggctttggct gatttggatc tggttgctga
     5881 acttccaaca attcctcaaa atcatccaca aagaactcct tcagtggagg gcgctttggc
     5941 ctcttcaggg tgttcagaag ctgctggatt ttggaggaca ctctgctgtt cacagggaca
     6001 ccatctgctg tttccagcat gctcccgctg cacccccgag gaacacttct cacagaagcc
     6061 acctgtggac gctcaaagta cgaatgctcc acgaggcccg tggtgacatc aggaggggca
     6121 gaatgcagat ctatgtgggt gtgggcctcg aggcctgcga gtgcagtggt agcggcggcc
     6181 cccggcgtgg ctgcagcact gggagcagtg gtgggtctcc ctcccgggtg agatgaggtg
     6241 gaggatgcag aggatgaggt ggacgagccc tgaatgaccc ggtcgagcca gggctcgacg
     6301 ctggaatggc tctggagcgg agtggaggtg agtcgcccgg gtcgccgtaa a
//
