LOCUS       sequence-491439-        7182 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  5' AAVLINK plasmid for TANC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7182)
  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  JOURNAL   Unpublished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 7182)
  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            "Unpublished"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: pAAVLINK-EF1alpha-HA-5'-TANC2
FEATURES             Location/Qualifiers
     source          1..7182
                     /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        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   4445..4585
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4445..4574
                     /label=AAV2 ITR
     rep_origin      4660..5115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4747..4766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4957..4978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5132..5151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5251..5273
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5311..5329)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5397..5501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5502..5570,5571..6362)
                     /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(5720..5739)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6533..7121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7022..7041
                     /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 gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatcttttc ggaatagtct caagatgctg cttactggtg ggaaatcaag
      781 tcgtaaaaac aggtcaagtg atggagggag cgaggaacca ccggatcgaa gacagtcaag
      841 tgtagactct cgccaaagcc gctctgggca aggtggcatc tccacagaaa gcgactgtgc
      901 ttttgagcca gactacgctg tcccgccact tccagtgagt gaaggtatgc agcacattcg
      961 gattatggag ggcatgtctc gctctcttcc atcctccccc ttactcacac atcagtctat
     1021 cagtgttcgg ctccagcctg tgaagaagtt aactgcccct ctgaggaagg caaagtttgt
     1081 tgagagccca cgaattccag aatccgagct tggctcacca accctcactt cagctcagaa
     1141 actggacgtg gatgcatact gccctggtga tgctgaacag gagcttggcc cccctccatc
     1201 tgtagatgag gcagcaaaca cactcatgac tcgtctgggt ttcctccttg gagagaaagt
     1261 cacagaagtt cagccaggtg accaatacag catggaagta caggatgaaa atcagacctc
     1321 agcaatcacc cagcggataa gtccttgttc cactctgact agcagcactg cctctccacc
     1381 agccagtagc ccctgctcta cactcccacc catcagtaca aatgcaactg ccaaggactg
     1441 cagctatggg gctgttacta gtccaacctc tacccttgaa agcagagata gtggcatcat
     1501 tgctacatta acaagctatt ctgaaaatgt ggaacgcaca aaatatgctg gggaaagcag
     1561 taaagaatta ggatctggag gaaacataaa accttggcaa tctcaaaaat ccagcatgga
     1621 ctcctgtttg tatcgagtag atgaaaacat gactgcttcc acctatagtc tgaataagat
     1681 cccagagaga aatttggaaa cagtgttatc tcaatcagta cagtctattc ctttgtatct
     1741 catgccacgg ccaaattcag tagcagccac cagctctgcc cacttggaag accttgctta
     1801 cctggatgag cagagacata cacctttgag aacttctcta cgaatgccaa gacagagcat
     1861 gggtggtgct cgcacacagc aggatttaag agttcgcttt gcaccttata ggcctccaga
     1921 tatctccttg aagcctctgc tctttgaagt gcccagcatc actacagagt cagtgtttgt
     1981 tggccgagat tgggttttcc acgaaataga tgctcaactt caaagttcaa atgccagcgt
     2041 gaaccaagga gtagtgattg tgggaaacat tggattcggc aaaactgcca tcatctccag
     2101 actggtggcc ctcagctgcc atggtacaag gatgagacag atcgcctcag acagcccaca
     2161 tgcctccccc aaacatgtgg atgccaacag agagctgcca ctcacacagc caccttcagc
     2221 ccactcatct atcaccagtg gaagctgccc aggaactccg gaaatgcgca ggcggcagga
     2281 ggaggctatg cgaagactag cctcgcaggt ggttgcctat cactattgtc aagcagataa
     2341 tgcctacact tgcttggtgc cagaatttgt ccacaatgtt gctgccttgc tctgccgctc
     2401 acctcagctg acagcctatc gggagcagct tcttcgggaa cctcacctgc agagcatgct
     2461 gagccttcgt tcctgtgttc aagaccccat ggcctccttc cggaggggag ttctggagcc
     2521 actagaaaat ctccataaag agagaaaaat cccagatgaa gatttcatca ttttaattga
     2581 tggattaaat gaagcagaat ttcacaaacc ggattatggg gatacaattg tatcgtttct
     2641 gagtaaaatg atcggaaagt ttccttcttg gctcaaacta attgtaacag ttaggaccag
     2701 tttacaggaa attaccaagc tgctgccttt ccataggatt tttttggatc gactagaaga
     2761 gaatgaagcc atagaccagg acctgcaggc ttacatcctg caccggatac acagcagctc
     2821 agagatccag aataacattt cacttaatgg caaaatggac aatactacat ttggcaaact
     2881 cagttctcat ctcaagaccc tcagtcaagg gtcctatcta tatctgaaac ttacatttga
     2941 cctcatagag aaaggctatc tagtgttaaa gagctctagc tacaaagtag ttcctgtttc
     3001 gctctcagag gtttatttac tccagtgcaa tatgaagttc ccaacccagt cttcctttga
     3061 ccgggtgatg cctctcctga atgtggcagt ggcctctctc cacccactga ctgatgagca
     3121 tatcttccag gccatcaatg ctgggagcat tgaaggcaca ctagaatggg aggattttca
     3181 gcagagaatg gagaacctct ccatgttcct aatcaagcgc agagacatga ctcgtatgtt
     3241 tgtacatcct tcttttcgag aatggcttat ctggagagaa gaaggagaga aaaccaaatt
     3301 tctctgtgat ccgaggagtg gtcacacgtt acttgccttc tggttttccc gccaagaggg
     3361 aaaactaaac cgacagcaga ctattgaact gggacatcac atcctcaaag cacacatttt
     3421 taagggtttg agtaaaaaag ttggtgtatc atcctccatc ctccaaggtc tctggatctc
     3481 ttatagcaca gaaggtcttt ccatggcact ggcgtcttta cgaaatctct acactccaaa
     3541 tataaaggtc agccgactgc tgattttggg aggtgccaat attaattacc ggacagaggt
     3601 tttaaataat gctccaattc tatgtgttca gtcccatctt ggttacacag aaatggtagc
     3661 cctgctgctg gagttcgggg ccaacgtgga tgcctcttct gaaagtggcc tgactccctt
     3721 gggatatgct gcagcagcag ggtacctgag cattgtggtg ctgctgtgca agaaacgggc
     3781 caaggtggat catttggata agaacgggca gtgtgctttg gttcatgctg cactccgagg
     3841 tcatctggag gttgtcaagt ttttgattca gtgtgactgg acgatggccg gccagcagca
     3901 aggagtattt aagaagagcc atgccatcca acaggccctc attgctgcag ccagcatggg
     3961 ttatactgag attgtctcct acctacttga tcttccagaa aaagatgaag aggaagtaga
     4021 gcgagcacag atcaacagct ttgacagtct ctggggagag acagccctaa cagctgcagc
     4081 cggaaggggc aaactggagg tgtgccgttt gctcttggaa caaggggcgg cagtggccca
     4141 gccaaaccgc cgaggagcag tgccactatt cagcacagtg cgccagggcc actggcagat
     4201 tgttgatctt ttactcaccc atggagctga tgtcaacatg gcagacaagc agggccgcac
     4261 tcccctgatg atggctgctt ccgaaggcca tctaggaacc gtggactttc tgcttgcaca
     4321 aggtaagtat caaggttaca agacaggttt aaggagacca ataagctatt attcgtatag
     4381 gatactttat acgaagttat taagttatgg agaccggtgt ggcacgtgcg gaccgagcgg
     4441 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4501 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4561 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4621 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4681 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4741 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4801 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4861 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4921 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4981 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5041 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5101 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5161 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5221 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5281 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5341 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5401 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5461 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5521 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5581 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5641 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5701 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5761 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5821 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5881 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5941 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6001 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6061 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6121 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6181 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6241 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6301 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6361 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6421 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6481 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6661 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6901 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6961 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7021 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7081 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7141 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
