LOCUS       sequence-491917-        7264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for TNRC6C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7264)
  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  JOURNAL   Unpublished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 7264)
  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            "Unpublished"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: pAAVLINK-EF1alpha-HA-5'-TNRC6C
FEATURES             Location/Qualifiers
     source          1..7264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3655..3672
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      4742..5197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4829..4848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5039..5060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5214..5233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5333..5355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5393..5411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5479..5583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5584..5652,5653..6444)
                     /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(5802..5821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6615..7203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7104..7123
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctga agaaaagaaa aagaaaaagc aggaagaaaa
      781 gaaaaagaag gaaggtgctc agaaaaaggc tgctgatcaa aaaaccaaag tgccagaacc
      841 tactaagacc tgttcaagcc agccccaacc tgccggtacc agtaccagta cttccaccag
      901 cactatctca agcagcaaca atggcaaacg tgcatctgcc agtggccagc agccagctgc
      961 atcccgttac ctgcctcgtg aggtgcctcc acgcttccgc cagcaagaac agaagcagct
     1021 attaaagaga ggccagccat tgcctacggg gactctaacc agtgtgagcc caacccaggg
     1081 tgctgggcct gcaggggtaa gcccacctcc cctacctgga gccggaacac agcatcatcc
     1141 cagtaagctc caaccagatc tcagtcatag tggattggca gatcattatg aaaattccca
     1201 ctggggacag cagcctactt acagaagcga agccaactgc agctgggata aagtgataat
     1261 agataggact gacaaggagg cgtggccttc catcacagga acagagactg aatctgcctc
     1321 agaatgtact acagacactg actctgcctc caactgtggc tcagagaaca gtagcatggc
     1381 tacagggagt gcccagggca acttcactgg acataccaag aagacaaatg gcaataatgg
     1441 caccaatggc gcactcgtcc aaagcccttc taatcagagt gcccttggag cagggggagc
     1501 gaacagtaat ggaagtgcgg ccagagtgtg gggtgtagcc acaggctcca gctctggtct
     1561 ggctcactgc tctgtcagtg gtggggatgg aaaaatggac actatgattg gagatgggag
     1621 aagtcagaat tgctggggtg cttccaactc caatgctggc attaatctta accttaatcc
     1681 taatgccaac ccagctgcct ggcctgtact tggacatgaa ggaaccgtgg cgacaggcaa
     1741 cccttccagt atttgcagtc cagtcagtgc cataggtcaa aatatgggca accagaacgg
     1801 gaacccaaca ggcactttag gtgcttgggg aaacttgctg ccacaagaga gcacagaacc
     1861 acaaacgtcc acttctcaga atgtgtcttt cagcgcacaa cctcagaacc ttaacactga
     1921 tggaccaaat aacactaacc ccatgaactc ttcacccaac cctatcaatg caatgcagac
     1981 aaatggactg ccaaactggg gcatggctgt tggtatgggg gccatcatcc cgccccacct
     2041 gcaaggcctt cctggtgcta atggatcatc agtttctcaa gtcagtgggg gcagtgctga
     2101 aggaataagc aattctgtgt ggggactgtc cccaggtaac cctgccacag gaaatagcaa
     2161 ttctgggttc agtcagggga atggagacac tgtgaactca gcattaagtg ctaaacaaaa
     2221 tggatccagc agtgctgtgc aaaaggaagg aagtggagga aatgcttggg attcaggacc
     2281 tcctgctggt cctggaatac tcgcctgggg aaggggcagt ggcaacaatg gcgttggtaa
     2341 tatccattca ggagcttggg gccaccccag ccgaagcacc tctaacggtg tgaatgggga
     2401 atggggaaag cccccaaacc agcattccaa cagtgacatc aatgggaaag gatcaacagg
     2461 gtgggagagt cctagtgtca ccagccagaa ccctaccgta cagcctggtg gtgaacacat
     2521 gaactcctgg gccaaagcgg catcttctgg aactacagca agtgaaggaa gtagtgatgg
     2581 ttctggcaac cacaatgaag gaagcactgg gagggaagga acgggagaag gccgaaggcg
     2641 agataaaggg attatagacc aagggcacat ccagttgcca aggaatgatc ttgacccaag
     2701 agttctgtct aatactggtt ggggacagac tcctgtaaag caaaacactg cctgggaatt
     2761 tgaagaatcc cctaggtctg aaaggaaaaa tgacaatggg acagaggcct ggggttgtgc
     2821 agctactcag gcttcaaact caggggggaa gaacgatggg tccatcatga acagtacaaa
     2881 tacctcttca gtatctgggt gggtcaacgc gccacctgcc gctgtgccag caaacacagg
     2941 ttggggagac agcaacaaca aagcgccaag tggcccgggg gtttgggggg actcgataag
     3001 ctctactgct gttagtactg ctgctgctgc caagagtggc catgcttgga gtggggccgc
     3061 aaatcaggag gacaagtcac ccacctgggg tgagcctcca aagcccaaat cccaacactg
     3121 gggagatgga caaagatcaa atccagcctg gagtgcagga gggggagatt gggcagattc
     3181 atcgtctgtc cttggacact tgggggatgg gaaaaaaaat ggatctggat gggatgctga
     3241 cagtaatagg tcagggtctg gttggaatga caccacgaga tctgggaaca gtggctgggg
     3301 caacagcaca aatacaaagg ccaatccagg tacaaactgg ggggagactt taaaacctgg
     3361 cccccaacag aactgggcta gcaaacccca agacaacaat gtgagtaact ggggaggagc
     3421 tgcttctgtg aaacagacag gaacagggtg gatcgggggg ccggtaccgg tcaaacagaa
     3481 ggacagcagt gaagcaactg gctgggaaga accctctcca ccgtccattc gccgcaaaat
     3541 ggaaattgat gatggtacct cagcttgggg ggacccaagc aactataaca ataaaactgt
     3601 aaacatgtgg gatagaaaca acccggtcat ccagagcagt accacgacca ataccaccac
     3661 caccaccacc actaccacga gcaacaccac acacagggtc gagacgccgc ccccgcacca
     3721 ggccggtact cagctgaatc gatcaccgtt gcttggtcca gtttcatcag gctggggaga
     3781 aatgcctaat gttcactcaa agactgaaaa ctcttgggga gaaccatcct ccccttctac
     3841 cctggtggat aatggcacag cagcatgggg gaagccaccc agcagtggca gcgggtgggg
     3901 agatcaccct gcagagccgc cggtggcatt tggaagagct ggcgcacctg ttgctgcctc
     3961 agccctgtgc aaaccagctt caaaatctat gcaagaaggc tggggcagtg gtggggatga
     4021 aatgaacctc agtaccagcc agtgggagga tgaagaaggg gacgtgtgga ataatgctgc
     4081 ttcccaagaa agcacctcct cctgcagctc ctgggggaac gcccccaaaa aaggacttca
     4141 aaagggcatg aagacgtctg gcaagcagga tgaggcctgg atcatgagcc ggctgatcaa
     4201 acaactcaca gacatgggct tcccgagaga gccagctgag gaggccttga agagtaacaa
     4261 tatgaatctt gatcaggcca tgagcgctct gctggaaaag aaggtggacg tggacaagcg
     4321 tgggctggga gtgaccgacc ataatggaat ggccgccaag cccctcggct gccgcccgcc
     4381 aatctccaaa gagtcttccg tggaccgccc cacctttctt gacaaggtaa gtatcaaggt
     4441 tacaagacag gtttaaggag accaataagc tattattcgt ataggatact ttatacgaag
     4501 ttattaagtt atggagaccg gtgtggcacg tgcggaccga gcggccgcag gaacccctag
     4561 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4621 aggtcgcccg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4681 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4801 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4921 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5041 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     5101 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     5161 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     5221 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5281 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5341 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5401 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5461 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5521 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5581 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5641 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5701 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5761 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5821 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5881 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5941 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6001 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6061 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6121 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6181 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6241 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6301 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6361 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6421 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6601 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6781 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7201 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7261 atgt
//
