LOCUS       sequence-476881-        5871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KDM2B expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5871)
  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  JOURNAL   Unpublished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 5871)
  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            "Unpublished"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' KDM2B
FEATURES             Location/Qualifiers
     source          1..5871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(198..224)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             261..529
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(542..753)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   798..927
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(989..1577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1069..1088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1748..2539,2540..2608))
                     /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     2371..2390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2609..2713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2781..2799
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2837..2859)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2959..2978
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2995..3450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3132..3153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3344..3363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3525..3665
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3536..3665
                     /label=AAV2 ITR
ORIGIN
        1 acgctgacga tctcctccac gtccgacaag tcctcgttct cgtcgtatcg ctggcggtca
       61 atcgggcgct gtgtggccga ctcaaattca aagcattttg tatatataac tgttttcttt
      121 ttttgctttt ctgctgcatg tctttttcgt ggggggtgat cctctgcaga tccccccatt
      181 tgcggacccg cagatccagc atagtcaggt acgtcataag ggtacatggt ggccgtacgg
      241 taggcgccgg tcacagcttg gatctgtaac ggcgcagaac agaaaacgaa acaaagacgt
      301 agagttgagc aagcagggtc aggcaaagcg tggagagccg gctgagtcta ggtaggctcc
      361 aagggagcgc cggacaaagg cccggtctcg acctgagctt taaacttacc tagacggcgg
      421 acgcagttca ggaggcacca caggcgggag gcggcagaac gcgactcaac cggcgtggat
      481 ggcggcctca ggtagggcgg cgggcgcgtg aaggagagat gcgagcccct cgaagcttca
      541 gctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta
      601 tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc
      661 agtttacccc gcgccacctt ctctaggcac ccgttcaatt gccgacccct ccccccaact
      721 tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag cgatcgcaga
      781 tccacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
      841 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
      901 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
      961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2761 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     2821 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2881 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2941 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     3001 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3061 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     3121 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3181 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     3241 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3301 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3361 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3421 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     3481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     3541 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     3601 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     3661 ttcctgcggc cgctcggtcc gcacgtgcca caccggtctc cataacttaa taacttcgta
     3721 taaagtatcc tatacgaata atagcttatt ggtctcctta aacctgtctt gtaaccttga
     3781 tacttacctt ccttgttgtc ccggttcatc ttctgctcct tgagcaggga gccgggcagg
     3841 ttggaggcgt acttaaagcc agggccacgc ttttgtttcc cggtcttgcc ggcgtggtta
     3901 cacttcggac actcccagca gtttggaagc tcgtcgttga ccacaccctc tgactcctta
     3961 atcttaaggc atccagggtg gatgatttca ttgcagatgg agcactccat gagcatgagg
     4021 ttaaacttgc cttcctcctc ttccaccgtg tcttccttcc ccgcctcgcc acacacaagg
     4081 cacacggcgg tgtggggcag cactggcgcg atgcactgcc gcatgatgca gctctgcttc
     4141 atgcgcccgg ggcccccgaa cttcttcatg tccttgcaga agtggcactc tccgcactcg
     4201 gtccgcaggc aggcctcgca cttgcggcat cgcgtccggc gccgccgagc tcctgccgtt
     4261 gtccggttgg cgaccaactt caccgcggag gccgggccca gcttcccctt gggccgaccc
     4321 acagcccggt tctttggagt cttctttggc caagtcacca cagggacccc agtgatggcc
     4381 agactagggt catcatctgc gtgctccttc aggacgttct tcacaccctc caggagtgcc
     4441 tgggggtcct cgatgccctc ggggacacac ttcttgttct ccgggaggga ttccagtttc
     4501 tccaccagag ctttcaggcc cttcagttca aactcagtga gatgggtcca tttggcagag
     4561 acctccgtgg cgggagagcc ggtgggggtc ttggggtagt ctacggccaa tgtggtggac
     4621 ttcacacttt cctccgactc attagacaaa gtccttttga ggaatcgcag ggcaggtgct
     4681 ttgggcttct tcccgagggc ctcctggtcc tcagagggcg tgctggtggg tgaagtggag
     4741 ccatcggtgg gcggtttggg tgccctgtcc ctgccctcgc cctcctcgtc cttctcctcc
     4801 tcctcctgag gctgctgatc acaggcctcc tcctccatct ccagccagga atccgaagag
     4861 aagccgtcta tgctgggctt cctcggggca tcaataagca tcgactccct ctggtattcc
     4921 tgagtgaggt gggagcgctg ggtcacacag tacacgtatc tctccaggac ataccagcac
     4981 atctcatagt agaaggggta acggaatttg ggctgcaccc gcgtcctgtc ctcgatctcg
     5041 tagatccgca gctgcatggg cacgttaaag ctgtgcagga tgtttccgcc gaacaccaaa
     5101 gagtctacag gggtgtagac ggcatggatc caaccggaag ggatgaaaaa tgtgtagccc
     5161 tgcttcagct caattctttg gcatcgttcc acacggtctc ccagaaagat gtcactctgt
     5221 ttgcctgaca gcacccactc ctcgtacagc gccaaattgt gcagcgttgg aggaatcagc
     5281 caaaaaatct tcccaccccg gaaaacatgg taccaaacgg aagtgcctcc aaagtcgatg
     5341 tggaagtcgg tgaaacaacc tttcacgctc atcagacagt actttttcac tttcgggtac
     5401 ttcatctctg caatggcgtt cgtggcttct gtctgcttct ccttcagatg ctggggccac
     5461 atgttgtcca cccagtccac caggtctacc acagtcggac gcttgaccaa gtgctccagc
     5521 ttggtgtggc tgaactctag gctgatgacg ttgtacagct tgtcccgctg ggcctcgggc
     5581 gtctcgtagt aacgcacaaa ctgggacatg ctcatctccg tgcccttctg ggtgttcaca
     5641 tccatcacgt ccacaagccg ccggctcccc actaggagtt tgacgtctcg gactgtgaaa
     5701 tcagggtcag gcatcttaat tcccagtcca tccttttctc gaaatatcag gggaaccctg
     5761 agagcttctc tctgtacgta ctcatagttg aaatctttgc cctccatggc gtgcacgaag
     5821 tccccctggt acagctggct gcgaagcttc tcctccaggc tgaagccgcg g
//
