LOCUS       sequence-476892-        6809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for AHDC1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  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  JOURNAL   Unpublished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 6809)
  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            "Unpublished"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-5' AHDC1
FEATURES             Location/Qualifiers
     source          1..6809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      51..506
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     348..369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(523..542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     642..664
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(702..720)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        788..892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(893..961,962..1753)
                     /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(1111..1130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1924..2512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2413..2432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2574..2703
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2748..2959
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2972..3240
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3277..3303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      5035..5044
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   6645..6785
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6645..6774
                     /label=AAV2 ITR
ORIGIN
        1 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
       61 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      121 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
      181 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
      241 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
      301 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
      361 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
      421 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
      481 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
      541 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      601 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      661 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
      721 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
      781 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      841 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      901 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      961 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1021 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1081 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1141 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1201 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1261 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1321 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1381 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1441 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1501 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1561 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1621 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1681 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1741 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1801 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1861 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1921 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1981 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2041 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2281 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2401 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2461 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2521 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     2581 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     2641 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2701 cctgcggccg cacgcgtgga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     2761 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     2821 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2881 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2941 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     3001 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     3061 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     3121 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     3181 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     3241 caagctgtga ccggcgccta ccgtacggcc accatgtacc cttatgacgt acctgactat
     3301 gctggatctc gtgtgaagcc ccagggcctg gtggtgactt ccagtgccgt gtgcagctct
     3361 cctgactacc tccgggaacc caagtactac cccggcggcc cccccacccc ccggcccctg
     3421 cttcccaccc ggccccctgc cagcccacct gacaaggcct tctccaccca cgccttctcc
     3481 gagaacccac gcccaccccc acgccgggac cccagcaccc ggcgcccacc agtccttgcc
     3541 aagggggacg acccgctgcc cccacgggca gcccgtcctg tctcacaggc ccgctgcccc
     3601 acaccggtcg gagacggcag cagctcccga cgctgctggg acaacgggcg ggtgaacctg
     3661 cgaccagtgg tgcagctgat tgacatcatg aaggacctga cacgcctctc ccaggacctg
     3721 cagcacagtg gtgtacacct ggactgtggt gggctccgac tgagccgccc gcctgcaccg
     3781 ccacccggcg acctacagta cagcttcttc tcctcaccca gtttggccaa cagcatccgt
     3841 agccctgagg agcgggccac cccacacgcc aagtcggagc ggcccagcca tcccctctac
     3901 gagcctgagc ctgagcctag ggacagtccc cagcctggcc aaggccatag tcccggagcc
     3961 acggctgcgg ccacgggtct gcccccagag cctgagccag acagcactga ttactcagaa
     4021 cttgctgacg ccgacatcct tagtgagctg gcctccctca cttgcccaga ggcccagctg
     4081 ctagaggccc aggccctcga gccaccatcg cccgagccgg agcctcagct cctggacccc
     4141 cagccccgct tcctggaccc gcaggcacta gagccgctcg gggaagctct ggagctgcca
     4201 cccctgcaac ctcttgctga tcctctgggg ctgccgggcc tggctctcca ggccctggac
     4261 accctgcctg actccttgga gtcgcagctg cttgaccccc aggcactcga ccccctgccc
     4321 aagctgcttg acgtcccagg tcgccgtctg gagccccagc agcccctggg gcactgccca
     4381 ctggccgagc ccttgcgcct ggacttgtgc tcaccgcacg gcccccccgg gcctgagggt
     4441 caccccaagt acgccttgcg gcgcactgat aggccaaaga tcctgtgtcg ccggcggaaa
     4501 gccggacggg gacgcaaggc agacgccgga cccgagggcc gcctactgcc cctgcctatg
     4561 cccacggggc tggtggctgc cctggccgaa cccccaccac caccgcctcc tccaccccct
     4621 gccctgccag gcccaggccc ggtctcagtc ccagagttga agccggaatc ttcccagacc
     4681 ccagtggtct ctacccgcaa aggcaagtgc cggggcgtgc ggcgcatggt ggtgaagatg
     4741 gccaagatcc ccgtatcgct ggggcggcgg aacaagacca catacaaagt gtcttccttg
     4801 agcagcagcc tgagcgtgga gggcaaggag ctgggcctgc gcgtgtcggc tgagcccacc
     4861 ccgctgctga agatgaagaa caatgggcgg aacgtggtag tggtcttccc acccggtgag
     4921 atgcccatta ttctcaaacg taagcgcggc cgccctccta agaacctgct gctgggtccc
     4981 ggcaagccca aggagccagc tgtggtggcg gccgaggcag ccactgtggc agcggccacc
     5041 atggccatgc cagaggtaaa aaaacgacgg cggcggaagc agaagctggc atctccccag
     5101 ccatcctatg cagcagacgc caacgacagc aaggccgagt actcagacgt cctggccaag
     5161 ctggccttcc tgaaccgcca gagccagtgc gctggacggt gctcaccgcc ccgctgctgg
     5221 acacccagtg agccggagtc ggtgcaccag gcccccgaca cccagagcat ctcccacttc
     5281 ctgcatcgtg tgcagggctt ccggcggcgt gggggcaaag caggcggttt tggtggccgg
     5341 ggtgggggcc atgcggccaa gtcagcccga tgctccttca gtgacttctt tgagggcatc
     5401 ggcaagaaaa agaaggtggt ggccgtggca gccgctgggg tcgggggccc tggccttact
     5461 gagttggggc acccacgcaa acggggccgg ggggaggtag acgctgtgac tgggaagcca
     5521 aagcgcaaga gacggtcccg gaagaatggg actctgttcc cagagcaggt gcccagtggc
     5581 ccaggctttg gggaggcagg tgctgagtgg gccggggata agggtggtgg ctgggcccct
     5641 caccatgggc acccaggcgg acaagctggc cgaaactgtg ggtttcaggg gaccgaggcc
     5701 cgggcctttg cctccactgg gctggagagt ggagcctcag gccgtggcag ctactacagc
     5761 acgggtgcac cctcaggcca gaccgagctc agccaggagc gccaaaacct cttcaccggc
     5821 tactttcgct cgctgctcga ttcggatgac tcctccgatc tcttggactt tgccctctca
     5881 gcctctcgcc cagagtcccg gaaggcatcg ggcacctatg cagggccacc caccagtgcc
     5941 ctgcctgccc agcggggcct ggccaccttc cctagccggg gagccaaggc cagcccagtg
     6001 gcagtgggta gcagcggggc tggggcggac ccctcctttc agcctgtcct gtccgcgcgc
     6061 cagaccttcc caccaggacg agcagcaagc tatgggctaa ctccagccac ttcagactgc
     6121 cgggcagccg agaccttccc caagctggtg cccccgccct cagccatggc ccgctcacct
     6181 accacccacc cgcctgccaa cacctacctg ccccagtacg gcggctatgg ggccggacaa
     6241 agcgtattcg ccccaactaa gccctttaca ggccaggact gcgctaacag caaggactgc
     6301 agcttcgcct atggcagtgg caacagcctc cctgcctcac ccagcagcgc ccacagcgcc
     6361 ggctatgccc caccgcctac cgggggcccc tgcctgccac caagcaaggc ctccttcttc
     6421 agcagctctg agggggcccc cttctctggt tcagccccca cgcccctgcg ctgtgacagc
     6481 cgggccagca cagtctcgcc cggtggctac atggtaccca aggtaagtat caaggttaca
     6541 agacaggttt aaggagacca ataagctatt attcgtatag gatactttat acgaagttat
     6601 taagttatgg agaccggtgt ggcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     6661 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     6721 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     6781 gcaggggcgc ctgatgcggt attttctcc
//
