LOCUS       sequence-491443-        7266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for PRR14L expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7266)
  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  JOURNAL   Unpublished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 7266)
  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            "Unpublished"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-EFS-HA-5'-PRR14L
FEATURES             Location/Qualifiers
     source          1..7266
                     /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        145..356
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             371..397
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      4744..5199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4831..4850)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5041..5062
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5216..5235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5335..5357
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5395..5413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5481..5585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5586..5654,5655..6446)
                     /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(5804..5823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6617..7205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7106..7125
                     /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 gcgtgggcag agcgcacatc gcccacagtc cccgagaagt
      181 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
      241 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      301 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagcgta
      361 cgccaccatg tacccttatg acgtacctga ctatgctgga tctctgtcat ctggagtaga
      421 gactcagcca gttccacttg attcctccat gtctgccgtg gtacaggaat tatactctga
      481 actcccagta agtgtctcca gagagcttca tgctgaccct gagccaagtg tgattccaga
      541 tgtaaaacct ggagcctcaa gctctctttt aagtcagaat agggcattgc ccttggagct
      601 gcagaggact catgtggaga gttgttgtga agaaacctat gagaccttgg atcatgggag
      661 tgagcctggg cgatgtgggc tagtggactc cacagcagga ggttctgtgg catctgggat
      721 cttggatagg gcaaagagaa gcgagagcat ggagccaaag gtcttcagag atccaggggg
      781 ccaggcagga attataagag aaccctctga gggagcaaag gaagatccac atcaacattc
      841 cacagctgct gaagaaaaga ctagcccgag tcaggaggat ctcctgatgc agtccagcaa
      901 agaactttca catgtggacc tccctgaaga ttttctaagg agcaaagaag gaaatgtaca
      961 gattacagct gaaactctgc taaaatccgc cgaagtacaa ggtatgaagg tcaatgggac
     1021 taagacggat aataatgaag gacacaaaaa tggcaatgtg agtaaagatc tctcagctgg
     1081 atgcggtgaa ttccaagaag tagacaaaat catgaccagt gatgaggttt cagaaaccag
     1141 cacattagtt accccagaac ctttaacctt tgtggaccct gtattaacag aagcaactcc
     1201 taaagaaaaa gaatgtgaag aattaaaaag ttgtccttgg ttgtcattac caggaaacag
     1261 tgccatttct aatgtggaca atgggaagga agagttgtgt aaaccaaacc tggtctgtga
     1321 agcagatgac aatcaccaac agcttcatgg ccaccataat gaacaaccca gttctacaca
     1381 tgatagtccc acagccacaa gccctttgaa agaaaattct gaagtttcat gtttcacatc
     1441 agacttgtct ggtccagaat ccagaacaat atccttagaa aactgtggtt ttgaaggtgg
     1501 tggtttgcta aagagatctg ctgaaaagac agacagttct tatttttata ggggggatga
     1561 tcaagggaag aacttggctt ctagagaaga aaatgaggaa cggcttttga ttcctaggag
     1621 tgaaagaggt ggaccttttc tttttaatgc cagggaacca gaaaaggaga ttagtggtcg
     1681 ttgttctggt gaaaaagagc ctgttgtttc tccaaaggaa aatatccaca ataactgcat
     1741 tcaagacagt ctccatacag ggaactctag ttctttaatg cccaattctt ttactgaagc
     1801 cacagaagta atgttaaata aaaatgattt gaaaattact gtacacgttc aaggtaactt
     1861 gacaaaccct gaggaccata aagaaacttt tactaatatg agccatcctg gtggacactc
     1921 tgaagaaagc agtttttcct ccttgatgca gattgaagag gcaggacaga caacccctgt
     1981 agagcccaat atattaagta aatcttttta cactaaagac tgtaactcct tagtcagcat
     2041 ccagagaaat ctggaaggca acacccagtt aaatgaagca tcatgtaatg attttctgtt
     2101 tgaaagaaaa tccattgtga gtttaatgcc agaggatcaa ataagtcctg taagtgaggt
     2161 attaaaaccc aagcaaggta ctgctctgtt gctaccatcc ccagaatttg attacagacc
     2221 tgagtcagaa aaagttatac agacctcaca tgatgatatt ccacttttag atgaacagag
     2281 catagcctgt gagatgaatg aactttcttg taccaatgaa ctggttgtaa acaaagtaga
     2341 aagtgaatgt gttttaaatc aacaagtgtc ccttaattct caagaacatg caaatttgcc
     2401 aactgactct ctactgcatt taaacaaaga gatgccttta gcaacaggca gagatgccca
     2461 tcagagccat caccctccat tagagggtag agcagatgtc attgctgata tacaaaccat
     2521 tcccattcag acaaaaataa aagacatctc tccaccaggt aaccaaacct gtggtgcctc
     2581 ttcaaactgt cccaccttaa acatcaaacc agtaagccta gagagaaaaa aggaaatggc
     2641 tgattcagga acaaaagctc tacattccag gctgcgctca aataagagag aagcagctgg
     2701 ctttcctcaa gtggtctctg tcatagaatg tcacagcgtt caatctcagg atatctctag
     2761 ctgtcatcgt gtaagaaaaa atgtatccca ggaaaacatg tgttctgctt ctgctgcttt
     2821 caagtccagc aaaatcagcc tgcaagttga taactctttg ataacaaaat atgaaaatgc
     2881 atttcagcac cgtgatcact gctgccaagg aacaggacac tctgtggaaa aaagcagctg
     2941 taaagtgagt tacacatcac aggaaaggga acttgatggg aaagaaacga atggcagcct
     3001 tccaggagat aagatcagaa acaaaatggt agcaggcttg ttaaatagtg gaatttcaaa
     3061 caaaaccatt cacacctcca gtagcatcaa actcagtgag gaagggctgg aaggaaagga
     3121 gcaggatgta tccaaagaaa ctgtattttg taagtataac atctctgatc atgctataca
     3181 agaactaaac cagactgtaa acattccagg tcctgaaaaa gtgttggacc agtctcctac
     3241 tgttatgttc tccagtttta aaaatgtaaa atcagttgaa acactcgatc agaaggcaga
     3301 tgaagtcctt gactgtcaga gtaaccaaaa cagaccagat gaatgcaaaa gtgaaggtca
     3361 gtcagccaag gagatgctaa gtagtgacca gagagagact gtcaccgagc ctcacgggga
     3421 ggtaaaccac aaccaaaagg atctgctggt cagctcaggc agtaataact cactaccttg
     3481 tggtagtcca aagaaatgca atttgaaagg agcctttgtc aagatgtctg gttgtgatga
     3541 gtccacagaa ggtatggtag acatcgtcta cacggactgt agtaataagc ttgcagaagg
     3601 tgtgctggac gtgaaggcat ctaatctact ggattgtggt gcaaggcaag agaaactggc
     3661 atttcaagag gactccagga gtaccttgtc tcggagagaa ctggatgctg cacatacagg
     3721 aacaactggt caggattcag atttcccagt tactgctgct tctacagtgg actttctcaa
     3781 aataaaaaaa tcatgtgaag aaaacgtatg cagatcttta aaagactgtg aaatggaaaa
     3841 gtgtccagac tcttgtgccc atgagatgga gtctgttgca gatcatgaac caaataaaag
     3901 aatattgggc agagtaaatt tgtctttaaa tgatagccat tatggacagc aagataaagg
     3961 aacatctctc agagaaacac aagaaatgac tgaaggatca agactagaac caaactctga
     4021 gtttggcaaa gaaagtacct ttggaatttc ctcaaaagag tcgatgtctt gccatgatga
     4081 aagctctgtt tccctaagaa gcctgaaatc cattgaaata atgccttctc aagaaaattc
     4141 agaaactaat gttaacagtg aagaaactga cctgaaaaat ctttgtaaac caaaagatgg
     4201 tgaaatgctt tgtgaaaatg taaaggactg cacagtcctt cctgagatga aggaaatagt
     4261 atcaagagat tggagtaatt ctagtgacag agacagcgta tgtacctgtg tggaaaagaa
     4321 tgcttgcaaa gcttgtcacc ctcacgagaa ttcctctgac agacatttgc ctttgacagt
     4381 gaaaacagat attaaagtga aaggagaaga aaccgaagag catcagaggt aagtatcaag
     4441 gttacaagac aggtttaagg agaccaataa gctattattc gtataggata ctttatacga
     4501 agttattaag ttatggagac cggtgtggca cgtgcggacc gagcggccgc aggaacccct
     4561 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4621 aaaggtcgcc cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4681 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4741 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4801 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4861 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4921 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4981 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5041 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     5101 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5161 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5221 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5281 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5341 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5401 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5761 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5821 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6001 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6121 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6421 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7261 acatgt
//
