LOCUS       sequence-491426-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for PHIP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  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  JOURNAL   Unpublished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 6811)
  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            "Unpublished"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'-PHIP
FEATURES             Location/Qualifiers
     source          1..6811
                     /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   4074..4214
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4074..4203
                     /label=AAV2 ITR
     rep_origin      4289..4744
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4376..4395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4586..4607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4761..4780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4880..4902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4940..4958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5026..5130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5131..5199,5200..5991)
                     /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(5349..5368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6162..6750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6651..6670
                     /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 ggatcttctt gtgagaggaa aggcctctcg gagctgcgat cggagctcta
      781 cttcctcatc gcccggttcc tggaagatgg accctgtcag caggcggctc aggtgctgat
      841 ccgcgaggtg gccgagaagg agctgctgcc ccggcgcacc gactggaccg ggaaggagca
      901 tcccaggacc taccagaatc tggtgaagta ttacagacac ttagcacctg atcacttgct
      961 gcaaatatgt catcgactag gacctcttct tgaacaagaa attcctcaaa gtgttcctgg
     1021 agtacaaact ttattaggag ctggaagaca gtctttacta cgcacaaata aaagctgcaa
     1081 gcatgttgtg tggaaaggat ctgctctggc tgcgttgcac tgtggaagac cacctgagtc
     1141 accagttaac tatggtagcc cacccagcat tgcggatact ctgttttcaa ggaagctgaa
     1201 tgggaaatac agacttgagc gacttgttcc aactgcagtg tatcagcaca tgaaaatgca
     1261 taaacgaatt cttggacact tgtcatctgt gtactgtgta acttttgatc gaactggcag
     1321 acggatattt actggttctg atgactgtct tgtgaaaata tgggcaacag atgatgggag
     1381 gttgttagct accttaagag gacatgctgc tgaaatatca gacatggctg taaactatga
     1441 gaataccatg atagcagctg gaagttgtga taaaatgatc cgagtctggt gtcttcgaac
     1501 ctgtgcacct ttggctgttc ttcagggcca tagtgcatct attacatcac tacagttctc
     1561 accattgtgc agtggctcaa agagatatct atcttctact ggggcagatg gcactatttg
     1621 tttttggctc tgggatgctg gaacccttaa aataaaccca agacctgcaa aatttacaga
     1681 gcgccctcgg cctggagttc aaatgatctg ttcttctttt agtgctggtg gaatgtttct
     1741 ggcgacggga agcacagatc atattattcg ggtttatttt tttggatcag gtcagccaga
     1801 gaaaatatca gaattggagt ttcatactga caaagttgac agtatccagt tttccaacac
     1861 tagtaacagg tttgtaagtg gcagtcgtga tgggacagca cgtatttggc aatttaaacg
     1921 aagagagtgg aagagcattt tgttggatat ggctactcgt ccagcaggcc aaaaccttca
     1981 aggaatagaa gataaaatca caaaaatgaa ggttactatg gtagcttggg atcgacatga
     2041 caatacagtt ataactgcag ttaataacat gactctgaaa gtttggaatt cttacactgg
     2101 tcaactaatt catgtcctga tgggtcatga agatgaggta tttgttcttg aaccacaccc
     2161 gttcgatcct agagttctct tttctgctgg tcatgatgga aacgtgatag tgtgggatct
     2221 ggcaagagga gtcaaaatac gatcttattt caatatgatt gaaggccaag gacatggcgc
     2281 agtatttgac tgcaaatgct ctcctgatgg tcagcatttt gcatgcacag actctcatgg
     2341 acatctttta atttttggct ttgggtccag tagcaaatat gacaagatag cagatcagat
     2401 gttctttcat agtgattatc ggccacttat tcgtgatgcc aacaattttg tattagatga
     2461 acagactcag caagcacctc atcttatgcc tccccctttt ttggttgatg ttgatggtaa
     2521 ccctcatcca tcaagatatc aaagattagt tcctggccgt gaaaattgca gggaggagca
     2581 actcatccct cagatgggag taacttcctc aggactgaat caagttttaa gtcagcaagc
     2641 aaaccaggag atcagcccac tggacagcat gattcaaaga ctacaacagg agcaagacct
     2701 gagacgttct ggtgaagcag gtatcagtaa taccagccgt ttaagtagag gctccataag
     2761 ttctacctca gaggttcatt caccaccaaa cgtaggacta agacgtagtg gacaaattga
     2821 aggtgtacgg caaatgcaca gcaacgcacc aagaagtgaa atagccacag agcgggatct
     2881 ggtagcttgg agtcgaaggg tggtagtacc cgagctatca gctggtgtag ccagtaggca
     2941 agaagaatgg agaactgcaa agggagaaga agaaataaag acttacaggt cagaagagaa
     3001 aagaaaacac ttaactgttc caaaagagaa taaaataccc actgtctcaa agaaccatgc
     3061 tcatgagcat ttcctggatc ttggagaatc caaaaagcaa cagacaaatc aacacaatta
     3121 tcgtacaaga tctgcattgg aagagactcc tagaccctca gaagagatag aaaatggcag
     3181 tagttcttca gatgaaggcg aagtagttgc tgtcagtggt ggaacatccg aagaagaaga
     3241 gagagcatgg cacagtgatg gcagttctag tgactactcc agtgattact ctgactggac
     3301 agcagatgca ggaattaatc tgcagccacc aaagaaagtt cctaagaata aaaccaagaa
     3361 agcagaaagc agttcagatg aagaagaaga atctgaaaaa cagaagcaaa aacagattaa
     3421 aaaggaaaag aaaaaagtaa atgaagaaaa agatggacca atatcaccaa agaaaaagaa
     3481 gcccaaagaa agaaaacaaa agagattggc tgtgggagaa ctaactgaaa atggtttgac
     3541 attagaagaa tggttgccat caacatggat tacagatacc attccccgaa gatgtccatt
     3601 tgtgccacag atgggtgatg aggtttatta tttccgacaa ggacatgaag cctatgtcga
     3661 aatggcccgg aaaaataaaa tatatagtat caatcccaaa aaacaaccat ggcataaaat
     3721 ggagctacgg gaacaagaac ttatgaaaat agttggcata aagtatgaag tgggattacc
     3781 taccctttgc tgccttaaac ttgcttttct agatcctgat actggtaaac tgactggcgg
     3841 atcatttacc atgaaatacc atgatatgcc tgacgttata gatttcctag tcttgagaca
     3901 acaatttgat gatgcaaaat acaggcgatg gaatataggt gaccgcttca ggtaagtatc
     3961 aaggttacaa gacaggttta aggagaccaa taagctatta ttcgtatagg atactttata
     4021 cgaagttatt aagttatgga gaccggtgtg gcacgtgcgg accgagcggc cgcaggaacc
     4081 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4141 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4201 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4261 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4321 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4381 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4441 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4501 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4561 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4621 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4681 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4741 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4801 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4861 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4921 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4981 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5041 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5101 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5161 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5221 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5281 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5341 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5401 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5461 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5521 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5581 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5641 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5701 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5761 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5821 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5881 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5941 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6001 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6301 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6781 ctggcctttt gctggccttt tgctcacatg t
//
