LOCUS       sequence-491757-        7237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for MED13L expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  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  JOURNAL   Unpublished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 7237)
  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            "Unpublished"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'-MED13L-FLAG
FEATURES             Location/Qualifiers
     source          1..7237
                     /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             3945..4010
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4087..4107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4500..4640
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4500..4629
                     /label=AAV2 ITR
     rep_origin      4715..5170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4802..4821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5012..5033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5187..5206)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5306..5328
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5366..5384)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5452..5556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5557..5625,5626..6417)
                     /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(5775..5794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6588..7176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7077..7096
                     /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 ggatttcctc accatctcgc
     1501 cattctcctt gccgttttgg gagaggctct tgttggaccc atatgggggc caccgtgatg
     1561 ttgcctatat tgtggtgtgt ccagaaaatg aggccttgct cgaaggagcc aaaactttct
     1621 tcagggactt gagtgctgta tacgagatgt gtaggcttgg gcagcacaag cccatctgca
     1681 aagtgctacg tgacgggatc atgcgcgtgg gaaaaactgt ggcacagaag ctgacagatg
     1741 agcttgtgag tgagtggttt aaccagcctt ggagcggcga ggagaatgac aatcattcca
     1801 gactcaaact ttatgcgcaa gtttgccgcc atcacctagc accttattta gccactctgc
     1861 agcttgatag cagcctattg ataccaccta aataccagac cccaccagca gcagcacagg
     1921 gacaagctac gccagggaat gctgggccct tagctccaaa tggatcagca gctcccccag
     1981 ctggcagtgc atttaatccc acctcgaata gtagttctac aaatcctgca gcaagtagtt
     2041 ctgcatctgg ttcctctgtg ccaccggtct catcgtctgc ctctgctcct ggtattagcc
     2101 agataagcac tacctcttct tcaggattca gtggtagtgt tggagggcag aaccccagca
     2161 ctgggggcat ttctgcggat agaacgcaag ggaacatagg ctgtggtgga gacactgacc
     2221 ctgggcagag ctcttctcag ccctcacagg atggacaaga gagtgttaca gaaagggaga
     2281 gaataggaat tcccacggag cctgactctg cagacagcca tgcccaccct ccagctgttg
     2341 tcatttacat ggtggacccg ttcacgtatg ctgcagagga ggactccact tctgggaact
     2401 tttggctgtt gagcttgatg cgctgctaca cagaaatgct ggataattta cctgagcata
     2461 tgagaaattc tttcattctc cagattgtgc cttgccagta catgctgcag acaatgaagg
     2521 atgagcaagt tttctacatt caatacttga agtccatggc attttcagtg tactgtcagt
     2581 gcaggcgacc actgcctaca cagatccaca ttaaatccct cacgggattt gggcctgcag
     2641 ccagcattga gatgaccctc aagaaccctg agcggcccag cccaatccag ctttactccc
     2701 ctccctttat attggcccca atcaaagaca agcagacaga gctgggagag acgtttggtg
     2761 aggcgagcca gaaatacaat gtgctcttcg tgggctattg tctgtctcac gaccagcgct
     2821 ggcttttggc ttcctgcact gacctccatg gggaattatt agagacctgc gttgtaaata
     2881 ttgctttacc aaacaggtca cggaggagta aagtatctgc acgtaaaatt ggactacaga
     2941 agttatggga gtggtgcata gggattgtcc aaatgacatc tctaccctgg agagttgtaa
     3001 tcgggcgact tgggcgtctt ggccatgggg agcttaaaga ttggagtatc ctccttggag
     3061 aatgttcact acagacaatc agcaaaaagc tcaaggatgt gtgccggatg tgtggaatct
     3121 ctgccgcaga ctctccttct atccttagtg cctgcctggt tgccatggag ccccaggggt
     3181 cctttgtagt gatgccagat gctgtcacaa tgggctctgt ttttggccga agtactgcac
     3241 tgaacatgca gtcatctcag ctcaacaccc ctcaagatgc ttcttgtaca cacatcttgg
     3301 tgttcccaac atcatcaacc atccaggtgg ctccagccaa ctaccccaat gaagatgggt
     3361 ttagccccaa caatgatgat atgtttgttg accttccatt cccagatgat atggacaatg
     3421 atattggcat attaatgact gggaacctcc attcctctcc caactcttcc ccagtaccct
     3481 ccccaggctc tccttctgga attggtgtgg gctctcactt ccagcatagt cggagccagg
     3541 gtgagcgtct tctttctaga gaagcaccag aggagctaaa gcagcagccc ctggcccttg
     3601 ggtattttgt atcaactgcc aaagctgaga atcttcccca gtggttttgg tcatcgtgtc
     3661 cccaggctca aaaccagtgc cctctcttct taaaggcttc gctgcatcac cacatttcag
     3721 tagcacagac agacgaactt ctgcctgcca ggaattctca gcgggttcca caccctcttg
     3781 actccaaaac cacgtcggat gttttaaggt ttgttttgga gcagtacaac gctctgtcct
     3841 ggctcacgtg caatccggcc acccaggacc gtacttcctg ccttcccgtc cactttgtgg
     3901 tgctcactca gttgtacaat gccatcatga atatactttg tacagactac aaagaccatg
     3961 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag taagaattct
     4021 gatgactcga gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4081 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4141 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     4201 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4261 cactgacaat tccgtggtgt ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4321 catctagctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4381 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     4441 tgtgggaggt tttttaaaga tctggagacc ggtgtggcac gtgcggaccg agcggccgca
     4501 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4561 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4621 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4681 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4921 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5041 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     5101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5161 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5221 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5281 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5341 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5401 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5461 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5521 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5581 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5641 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5701 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5761 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5821 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5881 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5941 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6001 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6061 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6121 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6181 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6241 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6301 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6361 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6421 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6721 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6961 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7201 cggttcctgg ccttttgctg gccttttgct cacatgt
//
