LOCUS       sequence-489074-        5773 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for IQSEC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  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  JOURNAL   Unpublished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 5773)
  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            "Unpublished"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'-IQSEC2
FEATURES             Location/Qualifiers
     source          1..5773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   159..299
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   170..299
                     /label=AAV2 ITR
     CDS             complement(2593..2619)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2656..2924
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2937..3148)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   3205..3334
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3396..3984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3476..3495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4155..4946,4947..5015))
                     /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     4778..4797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5016..5120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5188..5206
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5244..5266)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5366..5385
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5402..5773,1..84))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5539..5560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5751..5770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
       61 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc
      121 gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg
      181 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
      241 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      301 cggccgctcg gtccgcacgt gccacaccgg tctccataac ttaataactt cgtataaagt
      361 atcctatacg aataatagct tattggtctc cttaaacctg tcttgtaacc ttgatactta
      421 ccttagactg tcccgagaag aggagccgga gctgcagttg atggtctcat tggaattgct
      481 ggagctctca aggctctcgt tatctccacc atcagagttc tcgcctgctg cctcgcactt
      541 ccccaacctc cgacccccag ccacaccact gggcccagtg ccactgttgg gggctggtgg
      601 caggggccct ggtggggctg gggctgggcc ttcaggggct gggtagtggc ggtgtgggat
      661 cggggccctg cctggtggcc ccttgtgctt cagggtccca tgggggctgc agccatcagc
      721 ctcatacacc agctggcggt ggacagagcc gcgatctgag cggtcactca ggtccacgga
      781 gctgtcacta ggaggctcaa tggtaagcag gggaaggtgg gcagcccgca gccggaaatc
      841 ccggcactcc aagcacccgg gacccctgcg agtgccttcc tcacggctac cgtcttcccg
      901 ggttcctgat ggcactggtg gaggaactgg cgggagaggg gctggcgccc agaactcggg
      961 ccggccctgg ggtgggggtt ctgtgctggg cagtcgctca ggggattgtt ggggaactgt
     1021 gtcatccagg tagaggggaa gatcactgaa ggatgtggct gagctgtcct cttgctgttc
     1081 ctttgactcc cgcttctcca gctgggctga ccctggctcc tcagacatgg gccctgacgg
     1141 gtggcagttc agggcttcgt cgatggattc agccagagac tttacctgtt tggagaagga
     1201 gtcctcaagt tctgtgatgt cattagaaaa ctctccagat gtggtgacgg agcttccacc
     1261 accatcgatg cccccaccac aggagcctcc atatgggagc ccccgttcaa gccggtggct
     1321 ccgggcacca gccatggcac cctcgtccag cgaggcaggc ttgccctcga agtacgcggg
     1381 gttctgtgcc ttctcatact cctcaaagga gaactgcatc cgcatgttgg aaaggatgat
     1441 gcggcgggac atgcggctct ctgaggctga gctgcgtagc cgctcaaagt tcttgttcat
     1501 gcggtactgg cggaaggctg tctggatggt cctggcagcc ctgcggctca ggaaggagcc
     1561 cccatacttc ctctccagca tttccacctt cttgtcctgc aggtctgtgg agagttcata
     1621 gctgtccgat agggccttgg agcgctttat ctcctcctcc tcctgcttcc tcagggcgac
     1681 actggctggc tggaggcgtg cccgctgagc ccagggaagg cccactccag caggggggcc
     1741 ccccatgtgg ctgctggagg ggggcagctg gctcagccgg taggggggtt ggctcccagg
     1801 actatcaacc gctgtgctca ggtcactgcc tggggcatca ccctccacac tgactgttct
     1861 ggaattctca ccatcagatt ttctctggag ggtcgtggcc gggctggtgc tggtactggt
     1921 gctgtggcct ccgccggcgc cgggactgga gctcctggat gcgccacggc tcagctggcc
     1981 ccgctcccgc ggtggccgcg gccccacgcc caccgccgcc gaatagcccg cttccttctc
     2041 gcggcccggg tgcgccggcc cggcctcccg cccgccacgc tcgcgaccca gggctgggtt
     2101 ctcgtggtgc aggtggcact gggccacgtc acgctcgcgg gctgtgtaac cggtagtgtc
     2161 ctggagcggg taggaggcgt cccgctcctt gtcccgatac acagcctccc gattctggta
     2221 ggcgccttcc cggttcgggt agcccacgtc ccgggccgcc tggtggaact ggctctcccg
     2281 cagctcgcgg tggtggaact gggtctcgcg caggttctgg tactggctct cgcggcccgg
     2341 gctatcccgc gcgccgtggg ggtcccggtg cagctccccg cggtgcagct ggctctcctc
     2401 tcgcaggtcg cggttctcct gggtgagctg gtccagctgg ccctccagct cctcgatgcg
     2461 ccgccgctgg gtttccagca gctgctgctg gctctcgatg atgttgttca gctccagcag
     2521 gtactccacg gcccgatttg ggctctcgga tcccgggccg cccgggggcc ccgaccccgc
     2581 ctccatagat ccagcatagt caggtacgtc ataagggtac atggtggccg tacggtaggc
     2641 gccggtcaca gcttggatct gtaacggcgc agaacagaaa acgaaacaaa gacgtagagt
     2701 tgagcaagca gggtcaggca aagcgtggag agccggctga gtctaggtag gctccaaggg
     2761 agcgccggac aaaggcccgg tctcgacctg agctttaaac ttacctagac ggcggacgca
     2821 gttcaggagg caccacaggc gggaggcggc agaacgcgac tcaaccggcg tggatggcgg
     2881 cctcaggtag ggcggcgggc gcgtgaagga gagatgcgag cccctcgaag cttcagctgt
     2941 gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc acttatatac
     3001 ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact ttcccagttt
     3061 accccgcgcc accttctcta ggcacccgtt caattgccga cccctccccc caacttctcg
     3121 gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagcgatc gcagatcctc
     3181 tagagtcgac acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3241 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc
     3301 tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag
     3361 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3421 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3481 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3541 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3601 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3661 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3721 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3781 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3841 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3901 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3961 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4021 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4081 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4141 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4201 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4261 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4321 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4381 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4441 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4501 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4561 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4621 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4681 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4741 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4801 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4861 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4921 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4981 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5041 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5101 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5161 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     5221 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5281 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5341 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5401 aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5461 ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag
     5521 atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5581 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5641 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5701 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5761 gcgaaaggag cgg
//
