LOCUS       sequence-491399-        6215 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for RERE expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6215)
  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  JOURNAL   Unpublished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 6215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 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            "Unpublished"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'-RERE-FLAG
FEATURES             Location/Qualifiers
     source          1..6215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /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"
     CDS             2957..3022
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3087..3107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     rep_origin      3693..4148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3780..3799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3990..4011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4165..4184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4284..4306
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4344..4362)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4430..4534
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4535..4603,4604..5395)
                     /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(4753..4772)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5566..6154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6055..6074
                     /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 gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggaggcagca tagcgggggg
     1501 gtcgtcctgc ccactcccca ccgtccagat caaggaggag gctctggacg acgctgagga
     1561 gcctgagagc ccccctcccc caccaaggag cccgtccccg gagcccactg tggtggacac
     1621 ccccagtcac gccagccagt cagctaggtt ctacaaacac ctggaccggg gctacaactc
     1681 gtgtgcccgg acagacctgt acttcatgcc tctggccggg tccaagctgg ccaagaagag
     1741 ggaggaggcc attgagaagg ccaagcgcga ggctgagcag aaagcccgag aggagcgaga
     1801 gcgggagaag gagaaggaga aggagcggga gcgggagcga gagcgggagc gcgaggcaga
     1861 gcgggcggct aaggcgtcca gctcagcgca tgaaggtcgc ctcagtgacc cacagctcag
     1921 tggtcctggc cacatgcggc catccttcga gccaccacca accaccattg ctgctgtgcc
     1981 cccctacatc gggcccgaca cacctgccct tcggactctg agcgagtacg cccggcccca
     2041 cgtcatgtcg cccaccaacc gcaaccaccc cttctacatg ccccttaacc ccacggaccc
     2101 cctgctggcc taccacatgc ctggcctcta caacgtcgac cccaccatcc gcgagcggga
     2161 gctccgggag cgggagatcc gagagcggga gatccgagag cgggagctgc gggagaggat
     2221 gaagccgggc ttcgaggtga agcccccaga gctggacccc ctgcacccag ccgccaaccc
     2281 catggagcac tttgcccggc acagcgccct caccatcccc ccgaccgccg ggccccaccc
     2341 ttttgcttct ttccacccgg gcctgaaccc cttggagagg gagagactgg ccctggcggg
     2401 cccccagctg cggcccgaga tgagctaccc tgacagactg gcagccgagc gtatccacgc
     2461 agagcgcatg gcatcgctga ccagcgatcc cctggcccga ctgcagatgt tcaacgtgac
     2521 tccgcaccat caccagcact ctcacattca ctcccacctc cacctccacc agcaggaccc
     2581 cctccaccaa ggttcagcag gccccgttca cccgctggtc gaccccctga ctgccggtcc
     2641 ccacctggct cgcttcccct acccgcctgg cactctcccc aaccctctgc ttggacagcc
     2701 cccacacgag cacgagatgc ttcgccaccc agttttcggc accccctacc cccgtgacct
     2761 gcctggggcc atcccacccc ccatgtcagc agcccaccag ctgcaggcca tgcatgccca
     2821 gtcggccgag ctgcagagac tggccatgga gcagcagtgg ctgcatggac acccccacat
     2881 gcatggtggc cacctaccaa gtcaggaaga ttattacagt cgactgaaga aagaaggtga
     2941 caagcagtta tgtacagact acaaagacca tgacggtgat tataaagatc atgacatcga
     3001 ctacaaggat gacgatgaca agtaactcga gataatcaac ctctggatta caaaatttgt
     3061 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3121 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3181 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3241 acaggggctc ggctgttggg cactgacaat tccgtggtgt ttatttgtga aatttgtgat
     3301 gctattgctt tatttgtaac catctagctt tatttgtgaa atttgtgatg ctattgcttt
     3361 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3421 gtttcaggtt cagggggaga tgtgggaggt tttttaaaga tctggagacc ggtgtggcac
     3481 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3541 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc ggcctcagtg agcgagcgag
     3601 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3661 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3721 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3781 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3841 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3901 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3961 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4021 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4081 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4141 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4201 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4261 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4321 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4381 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4441 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4501 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4561 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4621 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4681 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4741 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4801 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4861 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4921 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4981 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5041 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5101 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5161 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5221 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5281 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5341 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5401 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5461 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5701 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6121 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6181 gttcctggcc ttttgctggc cttttgctca catgt
//
