LOCUS       sequence-491868-        5906 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for ATP10A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5906)
  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  JOURNAL   Unpublished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 5906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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'-ATP10A
FEATURES             Location/Qualifiers
     source          1..5906
                     /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        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   3169..3309
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3169..3298
                     /label=AAV2 ITR
     rep_origin      3384..3839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3471..3490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3681..3702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3856..3875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3975..3997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4035..4053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4121..4225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4226..4294,4295..5086)
                     /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(4444..4463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5257..5845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5746..5765
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctat ggagcgggag ccggcgggga ccgaggagcc
      781 cgggcctccg ggacggcgga ggcgccgaga gggcaggacg cgcacggtgc gctccaacct
      841 gctgccgccc ccgggcgccg aggaccctgc ggctggcgcg gccaagggcg agcggcgacg
      901 gcggcgcggg tgtgcccagc acctggccga caaccggctc aagactacca agtacacgct
      961 gctgtccttc ctgcccaaga acctgttcga gcagttccac cgcccggcca acgtgtactt
     1021 tgtcttcatc gcgctgctca acttcgtgcc ggcggtgaac gccttccagc ccggcctggc
     1081 actggcgccg gtgctcttca tcctggccat cacggccttc agggacctgt gggaggacta
     1141 cagccgccac cgctccgacc acaagatcaa ccacctgggc tgcctggtct tcagcaggga
     1201 agaaaagaaa tacgtgaacc gattctggaa agaaatccac gtgggagact ttgtgcgtct
     1261 tcgctgcaac gaaatcttcc ctgcggacat tctgctgctc tcctccagtg accccgacgg
     1321 gctatgccac atcgagaccg ccaacctgga tggagagacc aacctgaagc ggcggcaggt
     1381 ggtccgcggc ttctcggagc ttgtctccga attcaatcct ttgacgttca ccagcgtgat
     1441 cgaatgcgag aagccaaaca acgacctgag taggtttcgc ggctgcatca tacatgacaa
     1501 cgggaaaaag gccgggctgt ataaagaaaa cctgctgctg aggggctgca cccttaggaa
     1561 cacggacgca gtcgtcggca ttgtcatcta cgcaggacat gaaaccaagg ctctgctgaa
     1621 caacagtggg ccccgctaca agcgcagcaa gctggagagg cagatgaact gcgacgtgct
     1681 ctggtgtgtc ctgctccttg tttgcatgtc tctgttttca gcagtcggac atggactgtg
     1741 gatatggcgg tatcaagaga agaagtcatt attttatgtc cccaagtctg atggaagctc
     1801 cttatcccca gtcacagctg cagtttactc atttttaaca atgataatag ttctgcaggt
     1861 tttgatccca atttccttat acgtttccat tgaaattgtt aaagcatgcc aagtgtactt
     1921 cattaaccag gacatgcagt tgtatgacga agaaacagac tcgcagctgc agtgccgagc
     1981 tctgaacatc acggaagact taggacagat acagtacatt ttctcagata aaactggcac
     2041 tttgacagag aataagatgg ttttccgaag atgcactgtg tctggtgtag aatattctca
     2101 tgatgcaaat gcgcagcgtc tggccaggta ccaagaggca gactcggagg aggaggaggt
     2161 ggtgcccaga gggggctcgg tgtcccagcg cggcagcatc ggcagccacc agagtgtccg
     2221 ggtggtgcac agaacccaga gcaccaagtc ccaccggcgc acgggcagcc gggccgaggc
     2281 caagagggcc agcatgctgt ccaagcacac ggccttcagc agccccatgg agaaggatat
     2341 cacgcccgac ccaaagctgc tggagaaggt gagtgagtgt gacaagagcc tagccgtggc
     2401 gaggcatcag gagcacctgc tggcccacct ctcgcccgag ctgtctgacg tctttgattt
     2461 cttcatcgca ctcaccatct gcaacacagt cgtcgtcacg tccccggatc agccacgaac
     2521 aaaggtgagg gtgaggtttg agctgaagtc cccggtgaag acgatagaag acttcctgcg
     2581 gaggttcaca cccagctgcc tgacctcagg ctgcagcagc atcgggagcc tggccgccaa
     2641 caagtccagc cacaagttgg gctccagctt cccgtccacc ccgtccagcg acggcatgct
     2701 tctcaggctg gaggagaggc tgggccagcc cacctcggcc atcgccagca acggctacag
     2761 cagccaggcg gacaactggg cctcggagct tgctcaggag caggagtcag agcgcgagct
     2821 gcggtacgag gcggagagcc cggatgaggc cgcactggtg tatgcggcca gagcctacaa
     2881 ctgcgtgctt gtggagcggc tgcacgacca agtgtcagtg gagctgcccc acctgggcag
     2941 gctcaccttc gagctcctgc acacactggg tttcgattcc gtccgcaaga ggatgtcagt
     3001 ggtgatccgg cacccgctta ccgatgagat caacgtctac accaaggtaa gtatcaaggt
     3061 tacaagacag gtttaaggag accaataagc tattattcgt ataggatact ttatacgaag
     3121 ttattaagtt atggagaccg gtgtggcacg tgcggaccga gcggccgcag gaacccctag
     3181 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3241 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3301 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3361 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3421 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     3481 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3541 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3601 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3661 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3721 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3781 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     3841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3901 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3961 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4021 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4081 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4141 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4201 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4261 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4321 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4381 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4441 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4501 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4561 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4621 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4681 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4741 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4801 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4861 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4921 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4981 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5041 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5101 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5161 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5341 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5401 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5461 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5521 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5581 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5641 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5701 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5761 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5821 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5881 cttttgctgg ccttttgctc acatgt
//
