LOCUS       sequence-491390-        7067 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for IQSEC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7067)
  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  JOURNAL   Unpublished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 7067)
  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            "Unpublished"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'-IQSEC2-FLAG
FEATURES             Location/Qualifiers
     source          1..7067
                     /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             3031..3057
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             3787..3852
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3917..3937)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4330..4470
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4330..4459
                     /label=AAV2 ITR
     rep_origin      4545..5000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4632..4651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4842..4863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5017..5036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5136..5158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5196..5214)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5282..5386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5387..5455,5456..6247)
                     /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(5605..5624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6418..7006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6907..6926
                     /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 gggagcctcc ggctaccggc
     1501 ctgtgcaagc agacttacca gcgggagaca aggcatagct gggactcgcc agctttcaac
     1561 aatgatgtgg tccagaggcg gcactaccga atcggcctca acctcttcaa caagaagcca
     1621 gagaagggta tccagtatct gatcgagcgg ggcttcctgt cagacacacc ggtgggagtg
     1681 gctcacttca tcctggagcg gaaaggcctc agccggcaga tgatagggga attcctaggg
     1741 aaccggcaga agcagttcaa cagagacgtg ttggactgtg tggtggatga gatggacttc
     1801 tcctccatgg atctggatga tgcgctccgg aagttccagt cccatatccg ggttcagggt
     1861 gaggcccaga aagtggagcg actcatcgaa gccttcagcc agcggtactg tgtctgtaac
     1921 ccagccctcg tgcgccagtt ccggaaccca gacaccatct tcatccttgc ttttgccatc
     1981 atcctcctca ataccgacat gtacagtccc agcgtcaaag ctgaacgaaa gatgaaacta
     2041 gatgacttca tcaagaacct gagaggggtt gacaatggtg aagacatccc ccgagacctc
     2101 ctggtgggca tctaccagcg catccagggg cgtgaactgc ggaccaacga tgaccatgtg
     2161 tcccaggtgc aggctgtgga gcgcatgatt gttggaaaga aaccagtcct gtctctccct
     2221 caccgtcgac tggtttgctg ctgccagctc tacgaggtgc cagatccaaa ccgcccccag
     2281 aggctagggt tgcatcagcg ggaggtcttc ctcttcaatg atctccttgt ggtcaccaaa
     2341 attttccaga agaagaagat cttggtgacg tacagtttcc gtcagtcttt ccccctcgtg
     2401 gaaatgcaca tgcagctctt ccagaattca tattaccagt ttgggatcaa gttgctgtct
     2461 gcagtacctg gtggggagcg aaaagtcctc atcatcttca atgcccccag cctccaggac
     2521 cggctgcgct ttacatccga cctgcgcgag tccattgcgg aggtgcagga gatggagaaa
     2581 taccgtgtgg agtcggagct ggagaagcag aaaggtatga tgcggcctaa cgcctcacag
     2641 cctggagggg ccaaggactc agtgaatggg acgatggccc gcagtagcct ggaggacact
     2701 tacggggcag gcgatgggct caaacggggc gcactcagca gttccctgcg agacctctct
     2761 gatgcaggga agcgggggcg gcgtaacagc gtgggatcgc tggacagcac catcgaaggg
     2821 tctgttatta gcagtccacg ccctcaccag aggatgccac ctccgccccc accccctccg
     2881 ccagaggagt acaagagcca gaggcccgtc tccaactcct catccttcct gggctcccta
     2941 tttggaagca agcggggcaa ggggcccttc cagatgccac caccgccaac aggccaggcc
     3001 tctgcctcct cttcatctgc ttcttccacg caccaccacc accaccacca ccatcatggc
     3061 catagccacg gtggcctggg ggtgctgcct gatgggcagt ccaagctcca ggccctgcat
     3121 gcccagtatt gccaaggacc gggccctgcc ccgccaccct acctcccacc ccagcagccc
     3181 tctcttcccc cacctcccca gcagccccca cccttgcccc agctgggctc cattccaccg
     3241 cctcccgcct cagccccacc tgtggggcca catcgccact tccacgccca tggcccagtc
     3301 ccagggcccc aacactatac cttgggccgg ccaggcaggg cacccagacg gggggctgga
     3361 ggacaccctc agtttgctcc acatggccgc caccccctgc accagcccac atccccactg
     3421 cccctgtaca gtcctgcccc ccagcaccct ccagcccaca aacagggccc taagcacttc
     3481 atcttcagcc accacccaca gatgatgcca gcagcaggcg cggctggggg ccctggatcc
     3541 cggccaccag ggggctccta ctcccacccc caccaccccc agtcaccatt gtcaccacac
     3601 tcacccatcc caccccaccc ctcctatcca cccctccccc caccctcccc tcacaccccg
     3661 cactcacccc ttccacccac ctccccccat ggcccgctgc acgcctctgg gccccctggc
     3721 acagccaacc cccccagtgc aaaccccaag gccaagccaa gccggatcag caccgtggtc
     3781 tgtacagact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3841 gacgatgaca agtaactcga gataatcaac ctctggatta caaaatttgt gaaagattga
     3901 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3961 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4021 tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4081 ggctgttggg cactgacaat tccgtggtgt ttatttgtga aatttgtgat gctattgctt
     4141 tatttgtaac catctagctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4201 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4261 cagggggaga tgtgggaggt tttttaaaga tctggagacc ggtgtggcac gtgcggaccg
     4321 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     4381 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     4441 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     4501 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4921 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4981 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5041 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5101 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5161 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     5221 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5281 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5341 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5401 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5461 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5521 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5581 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5641 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5701 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5761 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5821 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5881 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5941 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6001 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6061 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6121 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6181 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6241 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6301 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6361 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
