LOCUS       sequence-491962-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for CUX1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  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  JOURNAL   Unpublished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 7319)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3'-CUX1-FLAG
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             273..1304
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1460..1479
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4039..4104
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4169..4189)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4582..4722
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4582..4711
                     /label=AAV2 ITR
     rep_origin      4797..5252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4884..4903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5094..5115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5269..5288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5388..5410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5448..5466)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5534..5638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5639..5707,5708..6499)
                     /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(5857..5876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6670..7258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7159..7178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt cgctctgcgc gaggcggctc cgccgcggcg gagggatacg
      181 gcgcaccata tatatatcgc ggggcgcaga ctcgcgctcc ggcagtggtg ctgggagtgt
      241 cgtggacgcc gtgccgttac tcgtacgcca ccatgtccaa tttactgacc gtacaccaaa
      301 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
      361 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
      421 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
      481 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa accatccagc
      541 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
      601 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
      661 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
      721 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
      781 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
      841 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
      901 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
      961 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     1021 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     1081 gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca
     1141 gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca
     1201 tgcaagctgg tggctggacc aatgtaaata ttgtaatgaa ctatatccgt aacctggata
     1261 gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga ttaggtaagt atcaaggtta
     1321 caagacaggt ttaaggagac caataactag tataacttcg tataggatac tttatagcaa
     1381 tttatgaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     1441 ggtcttactg acatccactt tgcctttctc tccacaggta acatcaccac ccggatccga
     1501 gcctcggaga ctggctctga tgaagccatc aagtccatcc tagagcaagc caagagggag
     1561 ctccaagtgc agaaaactgc agagccggcc cagccttcct ccgcatccgg cagcgggaac
     1621 tctgatgacg ccatccgctc catcctgcag caagcccgcc gggagatgga ggcccagcag
     1681 gctgccctcg accctgcctt aaagcaggca ccactgtccc agagtgacat caccatcctc
     1741 acccccaagc ttctgtccac ctcgcccatg cccaccgtgt ccagctaccc acctctcgcc
     1801 atctccctga agaagccctc cgcagctcct gaggccggtg cctctgctct gccgaacccc
     1861 ccggccctca aaaaggaggc ccaggacgcc cccgggctgg acccccaggg agcagccgat
     1921 tgtgcacaag gggtcctgag acaggtgaaa aatgaggtgg gccgcagcgg tgcctggaag
     1981 gaccactggt ggagcgcggt gcagccggag agaagaaatg ccgcctcctc cgaggaggcc
     2041 aaggccgaag aaacgggcgg cgggaaagag aagggcagcg gtggcagcgg aggtggcagc
     2101 cagcctcggg ccgagcgcag tcagctccag ggaccctcgt cgtcagagta ctggaaggag
     2161 tggcccagcg ctgagtcccc atactcccag agctcagagc tgagtctgac cggggccagc
     2221 cgcagcgaga caccacagaa cagccccctg ccatcctccc cgatcgtgcc catgtccaag
     2281 cccaccaagc cctcggtccc cccgctgacc cccgagcagt acgaggtcta catgtaccag
     2341 gaggtggaca ccatcgagct cacccggcag gttaaggaaa agctggccaa gaacggcatc
     2401 tgccagagaa tcttcgggga gaaggtgctg ggcctgtccc agggcagcgt cagcgacatg
     2461 ctgtcccgac cgaagccatg gagcaagctg acgcagaaag gccgagaacc cttcatccgg
     2521 atgcagctct ggctgaacgg cgagctaggc cagggtgttc tacccgtcca gggccagcag
     2581 caagggccag tcctccactc cgtgacatcg ctccaggacc cgctgcagca gggctgtgtg
     2641 agctcagaaa gcactccaaa gacctccgcc agctgcagcc ctgcccctga gtccccgatg
     2701 agttccagtg agtcggtgaa gagcctgacc gagctggtcc agcagccctg tccccccatc
     2761 gaggcgagca aggacagcaa gccaccagag cccagtgacc cgccagcatc cgactcccag
     2821 cccacaaccc cgctgcctct ctccggacac tcggccctca gcatccaaga attagtagcc
     2881 atgtccccgg agctggacac ctacggcata accaagcggg tgaaggaggt gctgacggac
     2941 aacaacctcg gccagcgctt atttggggag accatcttag ggctcaccca aggctctgtc
     3001 tctgacctcc ttgcccgccc caaaccctgg cataagctca gtctgaaagg acgagagccc
     3061 ttcgtccgga tgcagctgtg gctgaacgac cccaacaatg tggagaagct gatggacatg
     3121 aaacggatgg agaagaaagc ctacatgaag cggcggcaca gctcagtcag tgacagccag
     3181 ccctgcgaac cgccctctgt cggcaccgag tacagccagg gcgccagccc ccagccccag
     3241 caccagctga agaaaccccg ggtggtgctg gctccggagg agaaggaggc gctgaaacga
     3301 gcgtatcagc aaaagccata cccgtcacca aaaaccatcg aagacctcgc cacccagctc
     3361 aacctgaaaa ccagcaccgt catcaactgg ttccacaact acaggtctcg gatccgcaga
     3421 gaactgttca ttgaggaaat tcaggccggg agtcagggcc aggcgggcgc cagcgactca
     3481 ccctcggccc gcagcggccg ggcggcgccc agctcggagg gcgacagctg cgacggcgtg
     3541 gaggccactg agggcccagg cagcgccgac accgaggagc ccaagtctca gggagaggcc
     3601 gagcgggagg aggtgccgcg gccggcggag cagacggagc cgccgccctc ggggaccccg
     3661 ggcccggacg acgcccgcga cgacgaccac gagggaggcc ccgtggaagg cccggggccc
     3721 ctgcccagcc ccgcctccgc gaccgccacc gccgcgcccg cggcccccga ggacgccgct
     3781 acctcagccg ccgccgcgcc gggggagggc cccgcggccc cgagctccgc gccgccgccc
     3841 agcaacagca gcagcagcag cgccccccgc aggcccagct cgctgcagag ccttttcggc
     3901 ctccccgagg ccgcgggcgc ccgggactcg cgcgacaacc ccctgcgcaa gaagaaggcc
     3961 gcgaacttga acagcatcat ccaccgcctg gagaaggccg ccagccggga ggaacctatc
     4021 gaatgggagt tctgtacaga ctacaaagac catgacggtg attataaaga tcatgacatc
     4081 gactacaagg atgacgatga caagtaactc gagataatca acctctggat tacaaaattt
     4141 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4201 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4261 ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4321 ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt gaaatttgtg
     4381 atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga tgctattgct
     4441 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     4501 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gatctggaga ccggtgtggc
     4561 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4621 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4681 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4741 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4861 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4981 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5101 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     5161 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5221 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5281 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5341 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5401 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5461 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5521 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5581 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5641 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5701 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5761 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5821 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5881 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5941 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6001 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6061 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6121 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6181 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6241 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6301 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6361 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6421 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6481 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6541 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6601 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6781 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7261 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
