LOCUS       sequence-491441-        7410 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for TANC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  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  JOURNAL   Unpublished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 7410)
  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            "Unpublished"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'-TANC2-FLAG
FEATURES             Location/Qualifiers
     source          1..7410
                     /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"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4118..4183
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4260..4280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4673..4813
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4673..4802
                     /label=AAV2 ITR
     rep_origin      4888..5343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4975..4994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5185..5206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5360..5379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5479..5501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5539..5557)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5625..5729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5730..5798,5799..6590)
                     /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(5948..5967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6761..7349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7250..7269
                     /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 ggtgcctcca ttgctcttat
     1501 ggacaaagaa ggattgacag ccctcagctg ggcttgtttg aagggccatc tctcagtagt
     1561 acgttctctg gtggataacg gagctgccac agaccatgct gacaagaatg gccgtacccc
     1621 actggatctg gcagctttct atggcgatgc tgaggtggtc cagttcctgg tagatcatgg
     1681 ggccatgatc gagcacgttg actacagtgg aatgcgccct ttggataggg cagtggggtg
     1741 ccggaacact tctgttgttg tcactcttct gaagaaagga gccaagatag gttgtcagac
     1801 gttaccgagt cgcccacgag gtccagccac atgggcgatg gccacctcca agccagacat
     1861 catgatcatc ctgttgagca agctgatgga agagggggac atgttttata agaaaggtaa
     1921 agtaaaggaa gctgcccagc gctaccagta cgccctgaag aagttcccta gagaagggtt
     1981 tggtgaggac ttgaaaactt tccgggaact aaaggtgtct ctcctcctca acctctctcg
     2041 gtgtcgcagg aaaatgaacg attttggaat ggcggaggaa tttgctacta aggccctgga
     2101 gctgaaaccg aaatcttatg aagcttacta tgcgagagca agggcaaaac gcagcagcag
     2161 acagttcgca gcagccttag aggacctgaa cgaggccatc aagctgtgtc ccaacaaccg
     2221 tgagatccag agacttctgc tgagagtgga agaagagtgt agacagatgc agcagccaca
     2281 gcagccaccg ccgccaccgc agcctcagca gcagttgccg gaagaagcag aacctgagcc
     2341 acagcatgaa gacatatact ctgtacagga tatattcgag gaggagtacc tggaacagga
     2401 tgttgaaaat gtttccattg gcctccagac agaggcccgg cccagccagg ggctcccggt
     2461 catccagagc ccaccctcct ctcccccgca tcgggactca gcctacatct ccagctcacc
     2521 tcttggctct catcaggttt ttgacttccg gtccagtagt tctgtaggct ctcccactag
     2581 acagacctat cagtccacct cacctgccct ttctccaact catcagaact cacattacag
     2641 gcctagccca ccacacactt ccccggctca tcagggagga tcttaccgtt tcagcccccc
     2701 tcctgtggga ggacagggca aagaataccc aagccctccc ccttcccctc tccggagagg
     2761 ccctcagtat cgggccagcc ctccagctga aagtatgagt gtctatagat cccagtctgg
     2821 ttcacccgtg cgctatcagc aggaaacaag cgtcagtcag cttcctggca gacccaaatc
     2881 tccattatcc aaaatggccc agcggcccta ccagatgcct cagctccctg tggcagttcc
     2941 ccagcaaggg ctcaggctac agcctgccaa ggcccagatt gtgagaagta accagcccag
     3001 cccagccgtc cattcaagca ccgtcatccc cacaggagcc tatggccaag tagcccattc
     3061 aatggccagt aaataccagt cttcacaagg agacatagga gtcagccaga gccggttggt
     3121 ttatcaaggg tcaattgggg gaatcgtagg ggatggaagg ccggtgcagc atgtccaagc
     3181 cagcctgagt gcaggcgcca tctgtcagca tggaggattg accaaagagg atcttccaca
     3241 gcgaccttcc tcagcatacc gaggtggcgt gagatacagc cagacaccac agatcggacg
     3301 cagccagtca gcatcctatt acccagtctg tcactcaaaa ctagatctgg agcgctcctc
     3361 cagccaacta ggttcccctg atgtgtcgca tttaatcaga agacctatca gtgtcaaccc
     3421 taacgaaatc aaaccgcacc cgccaactcc caggccgttg ctgcattccc aaagtgtagg
     3481 ccttcgcttc tctccatcta gcaatagtat ctcctccacc tccaacctaa ctccgacctt
     3541 ccggccatct tcttccatcc agcaaatgga gatcccactg aaacctgcat atgagaggtc
     3601 atgtgacgag ctgtcgccag tgtctccaac tcaaggaggt taccccagtg agcccacccg
     3661 atccaggacc acaccattca tggggatcat agataaaaca gcacggactc agcagtaccc
     3721 ccacctccac cagcagaatc ggacctgggc agtgtcatct gtggacaccg tcctcagtcc
     3781 cacgtctcca ggcaacctgc ctcagcctga gtccttcagt ccaccatcat ccatcagcaa
     3841 cattgccttt tataacaaaa ccaacaatgc acagaatggc catttgctgg aggacgatta
     3901 ttacagcccc catgggatgc tggctaacgg gtctcgtgga gacctcttgg agcgagtcag
     3961 ccaggcctcc tcctatcccg acgtgaaggt agctcggact ctacctgtgg ctcaggcata
     4021 ccaggacaac ctgtacaggc agctgtcccg agactctcgg caagggcaga catcccctat
     4081 caaaccaaag agaccgttcg tggagtctaa tgttaaggac tacaaagacc atgacggtga
     4141 ttataaagat catgacatcg actacaagga tgacgatgac aagtaagaat tctgatgact
     4201 cgagataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4261 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4321 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     4381 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4441 aattccgtgg tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag
     4501 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4561 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4621 ggttttttaa agatctggag accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc
     4681 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4741 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4801 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4861 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4921 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4981 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5041 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5101 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5161 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     5221 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     5281 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     5341 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5401 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5461 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5521 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5581 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5641 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5701 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5761 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5821 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5881 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5941 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6001 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6061 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6121 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6181 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6241 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6301 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6361 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6421 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6481 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6541 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6601 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6661 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6721 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6781 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6841 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6901 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6961 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7021 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7081 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7141 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7201 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7261 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7321 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7381 tggccttttg ctggcctttt gctcacatgt
//
