LOCUS       sequence-491395-        7231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for MBD5 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  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  JOURNAL   Unpublished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 7231)
  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            "Unpublished"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-HA-5'-MBD5
FEATURES             Location/Qualifiers
     source          1..7231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      4709..5164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4796..4815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5006..5027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5181..5200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5300..5322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5360..5378)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5446..5550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5551..5619,5620..6411)
                     /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     complement(5769..5788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6582..7170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7071..7090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatctaatg gaggcaaaga gtgtgacgga ggggacaagg aaggaggtct
      781 tccagctata caagttcctg tgggttggca gcgtcgtgtg gatcaaaatg gagtgcttta
      841 tgtcagtccc agtgggtctt tgttatcttg cttggagcag gttaaaacat acctgcttac
      901 tgatggaaca tgcaagtgtg gcttggaatg tcctcttatt cttcccaagg tatttaattt
      961 tgatcctgga gctgctgtga aacagagaac cgcagaagat gttaaggcag atgaagatgt
     1021 cacaaagcta tgcatacata aaagaaaaat tattgcagtg gccacacttc ataaaagcat
     1081 ggaagcccca catccttctc tggtgctcac cagtcccgga ggaggaacaa atgcaactcc
     1141 agtagtacct tctcgggcag caactccaag atcagtaaga aataagtctc atgaaggaat
     1201 tacaaattct gtaatgcctg aatgtaagaa tcctttcaag ttaatgattg gatcatcaaa
     1261 tgccatggga aggctatatg tacaagaact gcctggaagc caacaacaag aactccaccc
     1321 tgtctacccc cgacagagat tgggcagcag tgaacatgga cagaaatctc cattccgtgg
     1381 cagccatgga ggcctgccca gcccagcgtc atcaggttcc cagatatatg gagatggttc
     1441 aatctctcca aggactgacc cacttggaag tcctgatgtt ttcacaagaa gtaatcctgg
     1501 ttttcatgga gctcccaatt ctagtcctat tcacctgaat aggactcctc tttctccacc
     1561 ttcagtaatg ctacatggtt ctcctgtaca gtcatcctgt gcaatggctg gaaggactaa
     1621 tatacctctt tccccaacct tgactacaaa gagtccagta atgaaaaaac caatgtgtaa
     1681 tttttcaact aatatggaaa taccacgagc aatgttccac cacaaaccac cccaaggccc
     1741 acctccccct cctccacctt cttgtgctct tcagaaaaag ccattaacat ctgagaaaga
     1801 tccacttggc attcttgacc ctattcctag taaaccagtg aatcagaacc ctgttatcat
     1861 taatccaacc agtttccatt caaatgtcca ctctcaggta cctatgatga atgtaagcat
     1921 gcctcctgct gttgttcctt tgccaagtaa tctcccattg ccaactgtaa aacctggtca
     1981 catgaatcat gggagtcatg tacaaagagt tcagcattca gcttcaacct ccctgtcccc
     2041 ttctccagtg acatcccccg tgcacatgat ggggactgga attggaagga ttgaggcatc
     2101 gccccaaaga tcacgctcat cttccacatc atcagatcat ggaaatttca tgatgccacc
     2161 tgtaggaccc caggccactt ctagtggtat taaggttcca cccaggtcac caaggtcaac
     2221 aatagggtcc ccaaggccat caatgccatc aagcccttct accaagtccg atggacatca
     2281 tcagtacaag gatatcccta acccattaat tgctggaata agtaatgtac taaatacccc
     2341 aagcagtgca gcttttccta ctgcatctgc cggaagtagt tctgtaaaga gtcagcctgg
     2401 tttgctggga atgcctttaa atcagatctt gaaccagcac aatgctgcct cctttccagc
     2461 aagtagttta ctctcagcag cagccaaagc acagctagca aatcaaaaca aacttgctgg
     2521 taacaacagt agcagcagta gcaattctgg agctgttgcc ggcagtggca acactgaagg
     2581 acatagcact ttaaacacca tgttccctcc tactgccaac atgcttctcc caacaggtga
     2641 agggcaaagt ggtcgagcag cactaagaga taagctgatg tctcagcaaa aagacgcatt
     2701 gcggaaaaga aaacaaccac ctacgacagt gttgagtttg ctcagacagt ctcaaatgga
     2761 tagttctgca gttcctaaac ctggacctga cttgctaagg aagcagggtc agggttcatt
     2821 tcccatcagt tcaatgtctc agttactaca gtctatgagt tgtcaaagct ctcacttgag
     2881 tagcaatagt accccgggtt gtggggcctc aaatactgct ttgccttgct ctgctaacca
     2941 gctgcatttt acagatccca gtatgaactc tagtgttctt cagaacatac ctttaagagg
     3001 ggaagccgtg cactgccaca atgcaaacac taactttgtt cacagtaaca gtccagtccc
     3061 caaccaccat cttgcaggtt taataaatca gattcaggct agcgggaact gtgggatgct
     3121 cagtcagtcg ggcatggctt taggaaattc cttacatccc aatccacctc agtcaagaat
     3181 ttcaacgtcc tccactccag tgataccaaa cagcattgtt agcagctata atcaaacaag
     3241 ttctgaagca ggcggttcag gaccatcatc ctccatagcc atagcgggca ccaaccaccc
     3301 tgccatcaca aagacaacat ctgttcttca agatggcgtc atagtcacca ctgcagctgg
     3361 aaacccactg cagagtcagc tacccattgg gagtgatttt ccttttgttg gccaggagca
     3421 cgcacttcat tttccatcca acagcacttc aaacaaccat cttccacacc ccttgaaccc
     3481 cagcctcctc agttctctac ctatctcttt gccagtgaat caacagcatc tcctaaacca
     3541 gaatctatta aatatcctcc agccttcagc aggagaaggc aagtctgaga tcaacctcca
     3601 ccctttaggt tttctcaacc cgaatgtaaa cgctgcttta gcttttctct ccagtgacat
     3661 ggatgggcag gtattgcagc ctgttcactt tcagctctta gcagccttgc ttcagaacca
     3721 agcccaagca gctgccatgc ttcccctgcc atctttcaat ctgaccatct cagatctttt
     3781 gcaacagcaa aatacccctt taccctcatt aacacagatg acagccccac cagaccattt
     3841 gccaagcaat cagtcagaca acagccgagc tgagaccctt ttaaccagcc ccctggggaa
     3901 ccctttacca agctttgcag gcagtgacac tacttttaac cccctgttcc tcccagctgt
     3961 caatggggcc tcaggattaa tgaccttgaa tccccagctg ttgggaggtg tcctgaactc
     4021 ggcatcggcc aacaccgcta atcatccaga ggtttccata gcaacctcct cccaggcaac
     4081 cactaccaca accactacat catcagcagt ggcagcactg actgtctcaa cacttggtgg
     4141 gacagcagtg gtgtcaatgg cagaaacatt gctgaatata tctaataatg ctgggaatac
     4201 acctggtcca gctaaactca acagtaactc tgtggtgcca cagctactta accctctact
     4261 ggggacaggt ctacttggtg atatgtcatc aataaacaat actttgagta accatcaact
     4321 gactcatcta cagtcgctgt taaacaacaa tcagatgttt cctccaaatc agcaacagca
     4381 gcaacttctc caggtaagta tcaaggttac aagacaggtt taaggagacc aataagctat
     4441 tattcgtata ggatacttta tacgaagtta ttaagttatg gagaccggtg tggcacgtgc
     4501 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4561 gctcgctcac tgaggccggg cgaccaaagg tcgcccggcc tcagtgagcg agcgagcgcg
     4621 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4681 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4741 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4801 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4861 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4921 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4981 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5041 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5101 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5161 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5221 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5281 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5341 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5401 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5461 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5521 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5581 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5641 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5701 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5761 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5821 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5881 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5941 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6001 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6061 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6121 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6181 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6241 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6301 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6361 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6421 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6481 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6541 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6601 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6661 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6721 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6781 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6841 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6901 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6961 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7021 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7081 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7141 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7201 ctggcctttt gctggccttt tgctcacatg t
//
