LOCUS       sequence-489085-        6537 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for KDM3B expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6537)
  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  JOURNAL   Unpublished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 6537)
  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            "Unpublished"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'-KDM3B
FEATURES             Location/Qualifiers
     source          1..6537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(531..557)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             594..862
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(875..1086)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   1131..1260
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1322..1910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1402..1421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2081..2872,2873..2941))
                     /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     2704..2723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2942..3046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3114..3132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3170..3192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3292..3311
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3328..3783)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3486)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3677..3696
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3858..3998
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3869..3998
                     /label=AAV2 ITR
ORIGIN
        1 ctcagtgcag catgttccaa aagaatctgg ttttggctat ctgttcccat ctgaaacaga
       61 tgcaacccat tggcatctga caggaaccga agctctcgat ccagaagata ttccactgga
      121 accacagaac ccaaacccag tttatctaca aggggagtaa aagtcagggc tggccattgt
      181 gcaatggaga ctcgaatgcc ctggagaagg actgggtcct cacggggcgc ccatacaaga
      241 gacccttcca gcagaagcac aataacttcc tcagcatgaa cttttaccca ggagtgttgc
      301 ttccagttac agccatcaaa ttctacaaag atcgctaggt cctggggcac cgccccgctc
      361 atggcccgca ccgtgccggc ccgccaggct cgagtgcgca gcgcaggccc cggatctccg
      421 ctcgccgctg ccgccgcggg agccgaggcc gaggctgagg ccgcagtgtc cgcgaacagc
      481 agcagcagcc gcttgcccac cggggaggcc gccgcgtccg ccatagatcc agcatagtca
      541 ggtacgtcat aagggtacat ggtggccgta cggtaggcgc cggtcacagc ttggatctgt
      601 aacggcgcag aacagaaaac gaaacaaaga cgtagagttg agcaagcagg gtcaggcaaa
      661 gcgtggagag ccggctgagt ctaggtaggc tccaagggag cgccggacaa aggcccggtc
      721 tcgacctgag ctttaaactt acctagacgg cggacgcagt tcaggaggca ccacaggcgg
      781 gaggcggcag aacgcgactc aaccggcgtg gatggcggcc tcaggtaggg cggcgggcgc
      841 gtgaaggaga gatgcgagcc cctcgaagct tcagctgtgt tctggcggca aacccgttgc
      901 gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa
      961 aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg
     1021 cacccgttca attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct
     1081 ctgcccactg acgggcaccg gagcgatcgc agatccacgc gtgcggccgc aggaacccct
     1141 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     1201 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3061 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3121 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3301 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
     3361 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
     3421 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
     3481 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3541 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     3601 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3661 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
     3721 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3781 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3841 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     3901 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     3961 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
     4021 ccacaccggt ctccataact taataacttc gtataaagta tcctatacga ataatagctt
     4081 attggtctcc ttaaacctgt cttgtaacct tgatacttac cttctgctag ggaggaagag
     4141 ggaatccaca ggttcatggc atcagggtca ctttgctggg ggcttaaaaa cccctccaca
     4201 cggagcaccc cttttcgagt gaagatcaac ctccggaagt gaaagaaacg gcaggctaca
     4261 gtagaatcat cttgctcctg ttccttaaac ttccggtacc gctccaggcg gcattcacga
     4321 cacttgtgca gatgaggtgc cacattgatg catgacccat cctgcaggaa gggctctcca
     4381 gattgcttca gcttcttcac tttgcttaca tctttcagca ctgactggcc aactttcagg
     4441 ggggcagtcc gaggccggcc cttgggggcc tgcttccctt tgcttttgcc cagcaacagc
     4501 tcagcactgc gctccccatt ggggcccagc ttccccatca ggtgctctgg aattcccttc
     4561 aggcctgtct tagcctgcag ctgctcctca gagtcactca aatctgacag gtcactgttg
     4621 gtgctggagt ccgagtcttg tctgcaagcc aagcttcgtt catccagtga gaaccttttc
     4681 acagcttcaa aaggagcttt gttctcctgt gaaaaatctg ccggggagga cagaaagccg
     4741 cctgagtgcc tgccaaagac gttactaaag gttttgagga ggggattgtc ctgctgccca
     4801 ggccccacag ttggcctctc ctctgtgggg ctagaggaga gagttggcat ctcaatgggc
     4861 tgagtgaggc tgctggtggg cgagctggag cggccattcc ccatggcaga gaggcttggg
     4921 cccccactgg taagagctga gcccagaaca ccagatacta gcttggagga gtcagttgta
     4981 atgaagaggg gtttcttctt tgctaaagat gccgaatctg cagaggagtg agactcgggc
     5041 cagctgggtg ccaccttgga ggtgacagtg gtagcagaag aggagctacc tgatgcctga
     5101 gatgcaaaac tactgaaagg gctgtgttca actttctcca caaatgccag gaaaggatta
     5161 gaaggctctt ctcgggacaa tttggggggc tgtaaaaata gattttcatg attctctggg
     5221 gtacgggcat ttaggaggct ccgtgggaag gctggagtga gggtgggcat ggactctgca
     5281 ggcactttcc ggtccacaga gctgccagcc ttagagcctg gcttagtgtc tgttccaaga
     5341 acggatctgc ctttacacag gcctgagctc aggctctcaa ggctttgttt gaatgagtcc
     5401 cgactagaag aatcatcagc caaactctct gaaacagtag tgaagtagtt actggtaggt
     5461 aaagtttggg acatgcattg aaaaaacaag tttttggaga gatcagtgtc tttctgtgcc
     5521 tcttgtgcag agctacagcc aaaagagaag cccaaaggtt tgttctctgt ggctagaaca
     5581 ccattggact ggctccttcc acttccaaaa gtcaaaggtt gcgatgaact agggagaggt
     5641 gctccaaatc cagaagaggc cagaatagaa tttcttgaat tctctcccga ggcctgcgac
     5701 cggctgcctt tctgtttttc tggcacagtc cctgctgcaa ggccagtgtc tgagatcctc
     5761 actgtgtttg gggtggagct ggcggtagtg accacagctg cggaagccac tatgccctgg
     5821 ccaacttgtt ccagtgtttt tcctgcctct ttgttttcgg ctccacccac ctctggggcc
     5881 aaaggtgtct gacctgtggc tgtagaatat ggagtgaaag atgcaggtgt gtccccacct
     5941 acaggctcat cctgcaccac cagagtcctg ccgttttctt tggtgtaagt ggcaaagcga
     6001 atgttgcggt ttatctgggg gacaaatgta gacggtggct gcttggctct ctgatccagc
     6061 cctggctctc cactggcatt ccctcctttc cagggccctc ggcttgcctc acctccatcg
     6121 ctcccattac tgtctacttc acccctgtct ccttttaatt tctttgatgc agggtcacac
     6181 ccagagtccg aagcactttt cctcctccgg ccatctttcc cctctatgct ttctctcttc
     6241 ttctttcctt tggaagattt tactgctttt aacgtacccc cctcgctttg aggcgttgaa
     6301 ttatccatca gcatcacatg gattagtctg ggatctaccg acttgatctc accactctca
     6361 gttacagaca cctccataag acgagtgatg gagtcctgat ggctcacaac accatagagc
     6421 cacccttctt ccccctctgg ctgatatatt ttgaccctgt gaccttgaac actgtaggga
     6481 cctcggctga atatctcttg tagcttttgg tcactgatca aagcattaac tgtttct
//
