LOCUS       sequence-491446-        7433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for MED13L expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7433)
  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  JOURNAL   Unpublished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 7433)
  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            "Unpublished"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'-MED13L
FEATURES             Location/Qualifiers
     source          1..7433
                     /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"
     repeat_region   4696..4836
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4696..4825
                     /label=AAV2 ITR
     rep_origin      4911..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4998..5017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5208..5229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5383..5402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5502..5524
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5562..5580)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5648..5752
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5753..5821,5822..6613)
                     /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(5971..5990)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6784..7372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7273..7292
                     /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 tctactgcgg cagcgaactg
      421 ggtggcgaac ggggcgagcc tggaggattg tcactccaac ctcttttcgc tggctgaact
      481 cacgggaatc aaatggcgta ggtacaattt tggagggcat ggggactgtg gacccataat
      541 ttcagcccca gcccaagatg atccaattct gttaagtttc atccgctgtc tgcaagctaa
      601 cctgctttgt gtatggcgtc gtgatgtcaa accagattgc aaagagttat ggatattctg
      661 gtggggagat gaacccaacc tagtgggtgt aatacatcat gaactgcagg ttgtggaaga
      721 aggactctgg gaaaatggcc tttcctatga atgtaggacg ctgctcttca aagcgatcca
      781 caatctgtta gaaaggtgcc taatggataa gaacttcgtt aggattggga aatggtttgt
      841 ccgaccctac gaaaaggatg aaaagccagt caacaaaagt gagcatttgt cctgtgcttt
      901 cacattcttt ctgcatggag aaagtaatgt atgcacaagt gtggagattg cccagcacca
      961 gccaatttat ttgatcaatg aggagcatat acacatggct cagtcttcac ctgcaccatt
     1021 tcaagtactg gtaagtcctt atggcttaaa tgggacgcta acaggccaag catacaagat
     1081 gtcagaccca gccactcgta agttgattga ggaatggcag tatttctacc cgatggtgct
     1141 aaaaaagaaa gaagaatcga aagaggaaga cgagttggga tatgatgatg atttccctgt
     1201 ggcagttgaa gtaattgttg gtggtgttcg gatggtttac ccttcagcat ttgttttgat
     1261 ctctcagaat gacatcccgg ttcctcagag tgttgccagt gctggaggcc acattgcagt
     1321 tgggcagcaa gggcttggta gtgtgaagga cccaagtaac tgtgggatgc ctctgacccc
     1381 tcccacctct ccagaacagg ctatcctagg tgagagtgga ggtatgcaga gtgctgccag
     1441 tcacctggtt tcccaagatg gagggatgat aacgatgcac agtccaaaga gatcggggaa
     1501 gattcctcca aaactccaca atcatatggt ccatcgagtc tggaaggaat gcatcctcaa
     1561 cagaacccag tccaagagga gccaaatgtc aactccaact cttgaagaag agcctgctag
     1621 caatcctgct acttgggatt ttgtggatcc aacccaaaga gtcagctgtt cttgttccag
     1681 gcataagctt ttaaaacgtt gtgcagtcgg gcccaatcga cctcccacag tatctcaacc
     1741 agggttcagt gcaggaccat catcatcttc atctttacca cctcctgctt cttctaagca
     1801 caaaacagca gaaagacagg aaaaaggaga caagctgcaa aagagaccct taataccatt
     1861 tcaccatagg ccctctgtgg ccgaagaatt atgcatggag caagatacac caggacagaa
     1921 actagggttg gcagggatag actcctcctt agaggtgtct agcagtagga aatatgataa
     1981 gcaaatggcc gtgccttcca gaaatacaag caagcaaatg aatctgaatc ctatggattc
     2041 acctcattcc cctatatccc ctctgccacc aacactcagc cctcagccac gaggtcagga
     2101 aacagagagt ttggacccac catcggtccc tgtgaatcca gccctttatg gaaatggact
     2161 agaactccag cagttgtcta ctctggatga cagaactgtc ctcgtaggcc aaagactgtc
     2221 tctcatggca gaggtcagcg agacagcctt atattgtggg attaggccct cgaacccgga
     2281 gtcatcagaa aagtggtggc atagttatcg tctcccaccc agtgatgatg ctgagttcag
     2341 gcctccagag ctccagggtg agagatgtga tgccaaaatg gaggtaaact cagagagcac
     2401 tgcattgcaa agactcttag cacaacctaa caaacggttt aaaatctggc aagacaaaca
     2461 gccccagttg cagccactcc acttccttga cccattgcct ctatcacaac aacctggaga
     2521 cagtttggga gaagtgaatg acccatatac ctttgaagat ggtgacataa aatacatctt
     2581 tacagccaac aagaaatgca aacaagggac ggagaaagat tccctgaaaa agaataagtc
     2641 agaggatgga tttggtacca aggatgtcac tacaccaggt cattccacgc cggtgcctga
     2701 tgggaaaaat gccatgtcta ttttcagttc tgctactaaa acagatgtcc ggcaggataa
     2761 tgctgctggc agagctggct ccagtagcct tacacaggta acagatttgg caccttccct
     2821 gcatgactta gacaacatct ttgataattc tgatgacgac gaacttgggg ctgtatcacc
     2881 tgctctgcgc tcatcaaaaa tgcctgcagt tgggacagaa gaccgacctc ttgggaagga
     2941 tggaagagct gctgttcctt atccaccaac agttgcagac ttgcaaagga tgtttcccac
     3001 tccaccatct ttggaacagc atcctgcatt ttctcctgtg atgaattata aagatgggat
     3061 cagctcagag acagtgacag cattaggcat gatggagagc cctatggtca gtatggtttc
     3121 aacacaactc acagaattca aaatggaagt ggaagatgga ttaggaagtc ccaagcccga
     3181 ggaaattaag gacttttcat atgtgcacaa agttccatcc tttcaacctt ttgtgggatc
     3241 ctccatgttt gctccactga agatgttgcc gagccattgt ttgctacctc tgaagatacc
     3301 tgatgcctgt ctgtttcggc cttcatgggc aattcctcct aaaattgaac aactgcccat
     3361 gccccctgca gccactttca ttagagatgg ctacaataac gtgcctagtg ttgggagcct
     3421 agcagatcca gactatctga acacaccaca gatgaacaca cccgtgacgt tgaacagcgc
     3481 tgccccagcc agcaatagtg gggcaggagt cctaccatct ccagcaaccc ctcgcttctc
     3541 tgtccccaca ccacgaaccc ccaggacccc aagaactccc agaggtgggg gcactgccag
     3601 tggtcaaggg tctgttaagt atgatagcac cgatcaagga tcaccagcct ccaccccctc
     3661 tactacacgg cccctcaact ctgtggagcc cgccaccatg cagccaattc ccgaagccca
     3721 cagcctctat gttaccctga ttctctccga ttccgtgatg aatatcttta aagacagaaa
     3781 ctttgacagc tgttgcatct gtgcctgcaa catgaacatc aaaggggcgg atgtcgggct
     3841 ttacatcccc gattcttcca atgaggacca gtaccgctgt acctgtgggt ttagtgcgat
     3901 tatgaaccgc aaacttggct acaattcagg actcttcctt gaagatgagt tggatatttt
     3961 tgggaagaat tctgatattg gtcaggctgc agagaggcgc ttaatgatgt gtcagtccac
     4021 cttccttcct caggtggaag gaaccaaaaa accccaggag ccacccataa gccttctcct
     4081 cctcctccag aatcaacaca cacaaccttt tgcttcactg aatttcctgg actacatttc
     4141 ctctaacaat cgccaaactc ttccctgtgt aagctggagt tatgaccggg tgcaagcaga
     4201 taataatgat tactggacgg aatgctttaa tgcgttggag caggggcggc agtatgtgga
     4261 taaccccact ggtggaaaag tggacgaagc tctggtgaga agtgccactg tgcactcttg
     4321 gcctcacagc aatgtgctgg acatcagcat gctctcctcc caggatgtgg ttcgtatgct
     4381 gttgtccctg cagccctttc tccaagatgc catccaaaag aagcgcacgg gcaggacctg
     4441 ggagaacatc cagcatgtgc agggaccact cacttggcag cagttccata aaatggcagg
     4501 acggggaacc tacggttcgg aagaatctcc tgagccgttg cccatcccca ctctgctggt
     4561 aggctatgac aaggtaagta tcaaggttac aagacaggtt taaggagacc aataagctat
     4621 tattcgtata ggatacttta tacgaagtta ttaagttatg gagaccggtg tggcacgtgc
     4681 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4741 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4801 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4861 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4981 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5101 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     5161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5221 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     5281 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5341 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5401 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5461 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5521 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5581 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5641 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5761 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5821 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5881 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5941 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6001 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6061 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6121 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6181 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6241 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6301 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6361 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6421 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6481 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6541 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6601 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
