LOCUS       sequence-489087-        7218 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for ARHGAP5 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7218)
  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  JOURNAL   Unpublished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 7218)
  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            "Unpublished"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'-ARHGAP5
FEATURES             Location/Qualifiers
     source          1..7218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2897..2923)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2960..3228
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3241..3452)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   3497..3626
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4447..5238,5239..5307))
                     /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     5070..5089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5308..5412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5480..5498
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5536..5558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5658..5677
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5694..6149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5831..5852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6043..6062
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6224..6364
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6235..6364
                     /label=AAV2 ITR
     RBS             7140..7148
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 aaacatcttc actttgatgg gttgattccc ttgtattatc agacaaataa gaattttcta
       61 tcatattttt tttctctaga acatcactaa aaaacagagt aaagacctca gtttgccgat
      121 gatactgaga taaagaatac agtgctgtaa atttagcagt gatctcagtt gcaatgtgtt
      181 ctccttcagt cattaactct ttgatagcct cattttcaaa aaaatctgct tggctgtcag
      241 taactgccac cagctgtaca ggaatggtgt cttgaacttc tgatagaaat gctcgaagca
      301 ttcccatcga agctttccgt tttgcagagt aaactaatat atacccatga actagttcat
      361 cttttcttac tccaattgaa gagtggtatg ataatattgt gatctgtatt cgcctccttt
      421 tttcaccaat gattttatca agcattaggg aattattctg tccagcttga gcagcactgc
      481 aagaatgaga atcaaggaag ggtgaaagaa taagatccac actaaatgga tctccacaca
      541 tggcgcacat gacaattctc aagtcagctt ctgataagtc cttattggca ggaattgggc
      601 tcaccacatc caaattgtgt ttaactgatt ccaatactcc tctgagagct tgctttattt
      661 gggtttcatt aaatttacga ggatatgtac cagcaggtac atctacaaaa ggacattgca
      721 acttgtttgc caactgctgc ccttggtgcc tgagaattgg tagattctta ctaatggaat
      781 ctctctgatt agccagaatt aatgtaaatg gaagattagc catgtattta tcttttctga
      841 tctgagaagc ttcagttctt attttcccaa taaattcccc aataaaactc aatgactcaa
      901 tggaattaaa tacacagaag cacccatgtg gtttaaaggc ggcagtccat aactgactca
      961 aaaagtaagg cgatttggca tcaaccggcc gaagatcaag ttcataaatt tttccatcta
     1021 aggcatactc atcatcagtg gattgtgtcc ttatctcatt tgctagttct tgggcaaggc
     1081 catccttccc taaaataaaa aggttaactt tatctatatt ggtactatcg tgatataatc
     1141 ttaagcggcc atgatccaac tgtaaaagac tactagcaag taactgctcc actttaatgt
     1201 ctgtacaatt ttggccacta agacatgttt ctttagtggg atgataaaca aatcctatat
     1261 gcttaagtag aagggattcc ctatcaggtg caagtttctg taaagcttta tatctaggtt
     1321 cttcactcag aactgtatga atttcactca ttttatctga actaggtgtt gcattaagat
     1381 ctaaatcata aaaaagttca gaatgctcaa aaagcatttc ttgaaactct tctttggctt
     1441 tttcaactat ttctcgctga tgcctaccat atacctcttt gctatcagcc tcagtgatat
     1501 atttgtaggc ttcatcctcc ataacaaagc acataacttc ctcccatggc tgccctggtg
     1561 aaatgaattg aattttttcc aaagtctttt tgaatttttc cttcatttcc accctcctct
     1621 tctcggatat cagatgctgt acatggttct gatagacttt ttcagcttct aaagtgctca
     1681 ggaggtcaaa tggaatccgc ctatcattaa ttttgtctat atggtcagtt tcatcccaag
     1741 gagttttttc tagcaccaca aaacataact ggaaatctgc tctcttttcc attaacttca
     1801 aagcttctga ccaattcaaa tgttcaatct cttctagatt tggcaaaaga gtgttaaaag
     1861 ctcttggtaa agtatttata tactcttctc tcctttttct tatatgttcc tgtttaagtt
     1921 gttctatatg ttttgagaat gtatttctgg cctttcttgt tccctctaag ttgatgtatt
     1981 cttcataatc aggatgattt tttaatttat tactaacagt tttccaagtt gcatgataat
     2041 ctctcacagt ctgcacaagt ttttcaaact tatctgttgc tgtgacaaca agttgtctct
     2101 gtgttttata agcatccaaa tagggaataa ttttaggctt gctacgagtt ttatccaaca
     2161 tttgtaccag tgcagtaaaa catgtttcaa tgttgacatt aaatcgtgct gatgtttcca
     2221 ctacaagaag gttcttttta tttgaagcaa atgcctgaac ttctctaaga taatgatcca
     2281 cgcattcatc acatttagtt gctgctatta ttacaggttt ttttgatttt gataactgga
     2341 caaaaaggtt attcacaaat ttaagttgat catcaaactt cctattgcat ccttgactta
     2401 catcaatgca taataaaaat ccatctacgt tgagcttccc ttcaggcatt tgcttctgtt
     2461 caaagtcttg ttctaagcct agctgatcag tgcaaatgta cattagtttt tctgctgact
     2521 gcaatttaga tgcagctgca cgttttatat atggttgcaa attcgtactc cgatgaggca
     2581 agaaagtctg gtcatcaatg aactctgttt gttcaatgac atgaattttg cattctactc
     2641 catcttcact attttgtatt atgtcacccc agtacaaaaa gtgatcattg tttactactc
     2701 gtcctccaaa gtcaatggtg ctaagcacag aagtatgctc tggataatat tcatctgctt
     2761 ttgagcgtac aaatctattg cacaaacaag actttccaac tccacagtta cctttgtctt
     2821 tttcagtccc agagagtcca actacactga tggtatagga tgggggacga ggctctttgt
     2881 tttttgccat agatccagca tagtcaggta cgtcataagg gtacatggtg gccgtacggt
     2941 aggcgccggt cacagcttgg atctgtaacg gcgcagaaca gaaaacgaaa caaagacgta
     3001 gagttgagca agcagggtca ggcaaagcgt ggagagccgg ctgagtctag gtaggctcca
     3061 agggagcgcc ggacaaaggc ccggtctcga cctgagcttt aaacttacct agacggcgga
     3121 cgcagttcag gaggcaccac aggcgggagg cggcagaacg cgactcaacc ggcgtggatg
     3181 gcggcctcag gtagggcggc gggcgcgtga aggagagatg cgagcccctc gaagcttcag
     3241 ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat
     3301 atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca
     3361 gtttaccccg cgccaccttc tctaggcacc cgttcaattg ccgacccctc cccccaactt
     3421 ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc gatcgcagat
     3481 ccacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3541 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
     3601 cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5461 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5521 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5581 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5641 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     5701 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5761 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     5821 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5881 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     5941 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6001 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6061 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6121 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
     6181 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc
     6241 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     6301 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     6361 tcctgcggcc gctcggtccg cacgtgccac accggtctcc ataacttaat aacttcgtat
     6421 aaagtatcct atacgaataa tagcttattg gtctccttaa acctgtcttg taaccttgat
     6481 acttaccttt ccaagtttca acaggagaaa gaagtggatc ttcttcactt ccacgatgtc
     6541 ttccttttct ttttgtttta gaagtggtga aagcctcatc atcactggca tctgaatgtg
     6601 ttcttctata gtatgacttg gctttactaa acaaggtttt agatttatat ttgtattttg
     6661 aaggcctccg ttccccatga ctttttgagg ttctatcaga aaacccattt tcttcatctc
     6721 cttgggtgtt atttacaaat gagtttcgaa ttttaatacg attttgacta tcatctggga
     6781 caacataaat ctcatcagaa tagcccttct gtttgaaaat tgtatcaatg ggttccgcat
     6841 agttatctga aggatccata tcttcaggat gtgccaaagg cacccgggaa gtctgctttc
     6901 ttggattttt accaatacca gcttcaattg tttttaaaag gtttggatcg agttttttca
     6961 catttgtctt aggtacaact ggtttaggtt taataggtgg aggcacttta tggttgcgtt
     7021 catggtcatg acagtttggg gtactatgaa taggatactc atttccttcc aaatctaatc
     7081 tatatctgga acggtcacta ggtgttggaa gcaactgtac atcatcccca attggactat
     7141 aaggaggtgg tgcttctgtg tcatcatctg aatcagggta gttattatag ggaaaacagt
     7201 ctctgggaga tggtagaa
//
