LOCUS       sequence-491866-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for CUX2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  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  JOURNAL   Unpublished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 6129)
  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            "Unpublished"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'-CUX2
FEATURES             Location/Qualifiers
     source          1..6129
                     /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   3392..3532
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3392..3521
                     /label=AAV2 ITR
     rep_origin      3607..4062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3694..3713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3904..3925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4079..4098)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4198..4220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4258..4276)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4344..4448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4449..4517,4518..5309)
                     /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(4667..4686)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5480..6068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5969..5988
                     /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 ggccgccaat gtgggatcga tgtttcaata
      781 ttggaagcga tttgatctac ggcgactcca gaaggagctt aattccgtcg cttctgagct
      841 gtctgcacgg caggaggaga gtgaacattc tcataaacat ttaattgaac tccgccggga
      901 atttaagaaa aatgtacctg aggaaatcag agagatggtg gctcctgtat taaaaagctt
      961 ccaagccgag gtggtggccc ttagtaagag aagtcaggag gcggaggctg cttttctgag
     1021 tgtttacaag caattaattg aagcaccaga ccccgtgcct gtgtttgagg cggcacgcag
     1081 cctagacgac agactgcagc cccccagctt tgaccccagt gggcagcccc ggcgagacct
     1141 ccacacttcg tggaagagga accccgagct cctcagcccc aaagagcaga gagaggggac
     1201 gtcgcctgcc gggcccacgc tgaccgaggg aagccgcctc ccaggcattc ccgggaaagc
     1261 cctcctgaca gaaaccttgc tgcagagaaa tgaggcggaa aaacaaaagg gccttcaaga
     1321 agtacagatc actttggcgg ccagactggg ggaggcagag gagaaaatca aagtcctaca
     1381 ttcagcgcta aaggctacgc aggcagagct gctagagctg cggcggaagt acgacgagga
     1441 ggcagcatcc aaggcagatg aagtcggcct gatcatgacc aacctggaga aagctaatca
     1501 gcgagctgag gctgcccagc gggaggtgga aagtctccgg gaacagctgg cctctgtcaa
     1561 cagctccatc cgcctggctt gctgctctcc ccaggggccc agtggggata aggtgaactt
     1621 cactctgtgc tcgggccctc ggctggaggc cgcgctggcc tccaaggaca gggagatcct
     1681 gcggctgctg aaggacgtgc agcacctcca gagctcactg caggagctgg aggaggcatc
     1741 cgccaaccag atcgccgacc tggagcggca gctcacggcc aagtccgagg ccatagaaaa
     1801 gctggaagag aagctccagg cccagtctga ctatgaggaa attaaaacgg agctgagcat
     1861 cctgaaagcc atgaagctgg cctccagcac ctgcagcctc ccccagggca tggccaagcc
     1921 tgaagactca ctgcttattg caaaggaggc cttcttcccc acgcagaaat tccttctgga
     1981 gaagcccagc ctcctggcca gccctgagga agacccatca gaggacgatt ccatcaagga
     2041 ttcactgggc acggagcagt cctacccctc ccctcagcag ctcccacctc caccagggcc
     2101 agaagacccc ctgtctccca gccctgggca gcccctgctg ggccccagct tggggcctga
     2161 cggcactcgg actttctcgc tgtccccctt ccccagcctg gcatcagggg agagactgat
     2221 gatgccccca gccgccttca agggagaggc gggcggcctg ctggtgttcc ccccagcctt
     2281 ctatggcgcc aagcccccca cagcccctgc caccccggcc cctggccctg agccactggg
     2341 cggtcctgag cccgcggatg gtggtggggg cggagcggcg gggcccgggg cagaggagga
     2401 gcagctggac acggcagaga tcgccttcca ggtgaaggag cagctgctga aacacaacat
     2461 cgggcagcgg gtgtttgggc attacgtgct ggggctgtcg cagggctcgg tcagcgagat
     2521 cctagcccgg cccaagccct ggcgcaagct cacggtgaag ggcaaggagc ccttcatcaa
     2581 gatgaagcag ttcctgtcgg atgagcagaa tgtactggcg ctcaggacca tccaagtgcg
     2641 gcagcgaggc agcatcaccc cgagaatccg cacgcctgag acaggctcag acgacgccat
     2701 caagagcatt ctagagcagg ccaagaagga gatcgagtcg cagaagggcg gcgagcccaa
     2761 gacctcggtg gccccgctga gcatcgccaa cggcacgacc cccgccagca cctcggagga
     2821 cgccatcaag agcatcctgg agcaggcacg ccgtgagatg caggcgcaac agcaggcgct
     2881 gctggagatg gaggtggcgc ccaggggccg ctcggtgccc ccctcgcccc cggagcggcc
     2941 atcactggcc actgcgagcc agaacggggc cccggccttg gtgaagcagg aggagggcag
     3001 cgggggcccc gcgcaggcgc cgctcccggt cctgtccccc gccgccttcg tgcagagcat
     3061 catccgcaag gtcaagtccg agatcggcga cgccggctac ttcgaccacc actgggcctc
     3121 cgaccgcggc ctgctcagcc gcccctacgc ctccgtgtcg ccctcgctgt cctcctcctc
     3181 ctcctctggc tactctggcc agcccaacgg ccgcgcctgg ccccgcgggg acgaggcccc
     3241 tgtgcccccc gaggacgagg cggcggcagg taagtatcaa ggttacaaga caggtttaag
     3301 gagaccaata agctattatt cgtataggat actttatacg aagttattaa gttatggaga
     3361 ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3421 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3481 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3541 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3601 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3661 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     3721 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3781 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3841 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3901 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     3961 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4021 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4081 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4141 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4201 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4261 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4321 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4381 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4441 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4501 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4561 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4621 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4681 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4741 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4801 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4861 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4921 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4981 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5041 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5101 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5161 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5221 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5281 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5341 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5401 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5461 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5581 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6121 ctcacatgt
//
