LOCUS       sequence-491872-        5531 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for CUX1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5531)
  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  JOURNAL   Unpublished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 5531)
  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            "Unpublished"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'-CUX1
FEATURES             Location/Qualifiers
     source          1..5531
                     /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      3009..3464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3096..3115)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3306..3327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3481..3500)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3600..3622
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3660..3678)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3746..3850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3851..3919,3920..4711)
                     /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(4069..4088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4882..5470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5371..5390
                     /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 ggatctttgt gcgtagccgg agccaggttg aagagagaac tcgatgccac
      781 cgcaacggta ttggcgaacc ggcaggatga aagtgagcag tccagaaagc ggcttatcga
      841 acagagccgg gagttcaaga agaacactcc agaggatttg cgcaagcagg tagcgccgct
      901 gctgaagagt ttccaaggag agattgatgc actgagtaaa agaagcaagg aagctgaagc
      961 agctttcttg aatgtctaca aaagattgat tgacgtccca gatcccgtac cagctttgga
     1021 tctcggacag caactccagc tcaaagtgca gcgcctgcac gatattgaaa cagagaacca
     1081 gaaacttagg gaaactctgg aagaatacaa caaggaattt gctgaagtga aaaatcaaga
     1141 ggttacgata aaagcactta aagagaaaat ccgagaatat gaacagacac tgaagaacca
     1201 agccgaaacc atagctcttg agaaggaaca gaagttacag aatgactttg cagaaaagga
     1261 gagaaagctg caggagacac agatgtccac cacctcaaag ctggaggaag ctgagcataa
     1321 ggttcagagc ctacaaacag ccctggaaaa aactcgaaca gaattatttg acctgaaaac
     1381 caaatacgat gaagaaacta ctgcaaaggc cgacgagatt gaaatgatca tgacggacct
     1441 tgaaagggca aaccagaggg cagaggtggc tcagagagag gcggagacct taagggaaca
     1501 gctctcatcg gccaatcact ccctccagct ggcctcacag atccagaagg caccagacgt
     1561 ggagcaggcc atagaggtgc tgacccgctc cagcctagaa gttgagttgg ccgccaagga
     1621 gcgggagatc gcacagctgg tggaggacgt gcagagactc caggccagcc tcaccaagct
     1681 gcgggagaat tcggccagcc agatctcaca gcttgagcag cagctgagcg ccaaaaacag
     1741 cacactcaaa caactggaag aaaaactcaa aggccaggct gactatgaag aggtgaagaa
     1801 agagctgaac attctgaagt ccatggagtt tgcaccgtcc gagggcgctg ggacacagga
     1861 tgcggccaag cccctggagg tgctgttgct ggagaagaac cgctcgctgc agtccgagaa
     1921 cgccgcgctg cgcatctcca acagcgacct gagcgggtca gccaggagga aagggaaaga
     1981 ccagcctgaa agtcggcgcc cgggatcttt gccggccccc cctccttctc agttgccccg
     2041 caacccgggg gagcaggctt ccaatactaa tggtacacac cagttctcac cagcggggtt
     2101 aagtcaagac tttttcagct catccctggc aagccccagc ctacccctgg cttctacagg
     2161 aaaatttgca ctaaactctc ttctccagcg gcagctaatg cagtccttct actccaaggc
     2221 tatgcaggaa gccggaagca caagcatgat tttttcaaca ggtccataca gcacaaactc
     2281 catatcttcc caaagtccat tacaacaaag cccagatgtc aatggcatgg ccccatcccc
     2341 cagccagtca gaaagtgctg ggagcgtctc cgagggcgag gagatggaca ctgcagaaat
     2401 cgcccggcag gtcaaagagc agctgattaa gcacaatatc ggacaacgta ttttcggaca
     2461 ttatgtgttg ggactgtctc aagggtccgt gagcgagatt ctggcccggc ccaagccatg
     2521 gaataaactg actgttcgtg gcaaggagcc atttcacaag atgaaacagt tcctctccga
     2581 tgagcagaac atcctggccc tccgtagcat ccaaggcaga caaagagaga atccaggcca
     2641 gagcctgaac agactatttc aggaagtacc gaaacgaaga aatgggtctg aaggtaagta
     2701 tcaaggttac aagacaggtt taaggagacc aataagctat tattcgtata ggatacttta
     2761 tacgaagtta ttaagttatg gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa
     2821 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2881 cgaccaaagg tcgcccggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2941 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3001 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3061 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3241 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3301 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     3361 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     3421 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     3481 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3541 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3601 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3661 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3721 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3781 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3841 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3901 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3961 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4021 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4081 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4141 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4201 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4261 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4321 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4381 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4441 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4501 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4561 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4621 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4681 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4741 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4801 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4861 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4921 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4981 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5041 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5101 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5161 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5221 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5281 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5341 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5401 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5461 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5521 tgctcacatg t
//
