LOCUS       sequence-476883-        6817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for PHRF1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  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  JOURNAL   Unpublished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 6817)
  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            "Unpublished"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' PHRF1
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(671..697)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             734..1002
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(1015..1226)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   1271..1400
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1462..2050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1542..1561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2221..3012,3013..3081))
                     /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     2844..2863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3082..3186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3254..3272
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3310..3332)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3432..3451
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3468..3923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3605..3626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3817..3836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3998..4138
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4009..4138
                     /label=AAV2 ITR
ORIGIN
        1 ccaccggcac ctcctggaga ggggggtcca agcattccat gtggtacccc gcatcacagc
       61 cgtcgcagag caaaagcctg tcctcacggt cgctcctgcc gcacacctca cagaaggtcg
      121 ggtcctcctc ctcctcgctc gctttggtgt tctccactgg gatctttctt aagattttac
      181 caccaaattg agctcgaata caaatgcact taaatagagt tcgatcaact ggacaggaat
      241 tggcattctt ggaccattcg acaatgcagt ccaggcagaa gtaatgggca cagttctccg
      301 gcgtccccac ggcctggtct ctgaatgcgt tgagacagat tgggcagctc tctgcatcat
      361 catcagaatt gaaagagcca gcggcttcca gtttcccctg agtacccgct acctccagca
      421 atgtctcccc gtcgtcttca gaatcctcgg aaccagatct gtcttccagg tcttcctcct
      481 cagacgcccc ctcgtcttct ccgtctgtgc catctccatg ctcgctgtca ctgtcgtccc
      541 cagagtcccc actgctgccc acgctgcttt cttcaaagtc acctgccggg tctgcagggc
      601 cgacctgtgg gtgtccatcc ggccctgggc tccgggccac aagctcatcc aggctgtcgt
      661 catcagatcc agcatagtca ggtacgtcat aagggtacat ggtggccgta cggtaggcgc
      721 cggtcacagc ttggatctgt aacggcgcag aacagaaaac gaaacaaaga cgtagagttg
      781 agcaagcagg gtcaggcaaa gcgtggagag ccggctgagt ctaggtaggc tccaagggag
      841 cgccggacaa aggcccggtc tcgacctgag ctttaaactt acctagacgg cggacgcagt
      901 tcaggaggca ccacaggcgg gaggcggcag aacgcgactc aaccggcgtg gatggcggcc
      961 tcaggtaggg cggcgggcgc gtgaaggaga gatgcgagcc cctcgaagct tcagctgtgt
     1021 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
     1081 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
     1141 cccgcgccac cttctctagg cacccgttca attgccgacc cctcccccca acttctcggg
     1201 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagcgatcgc agatccacgc
     1261 gtgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     1321 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg
     1381 agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2221 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2281 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2341 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2401 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2461 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2521 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2581 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2641 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2701 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2761 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2821 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2881 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2941 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3001 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3061 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3181 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3241 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     3301 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3361 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     3421 gcttaactat gcggcatcag agcagattgt actgagagtg caccataaaa ttgtaaacgt
     3481 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3541 gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag agttgagtgt
     3601 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3661 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     3721 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3781 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3841 cgctaaggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3901 taatgcgccg ctacagggcg cgtactatgg ttgctttgac gtatgcggtg tgaaataccg
     3961 cacagatgcg taaggagaaa ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc
     4021 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     4081 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     4141 ggccgctcgg tccgcacgtg ccacaccggt ctccataact taataacttc gtataaagta
     4201 tcctatacga ataatagctt attggtctcc ttaaacctgt cttgtaacct tgatacttac
     4261 ctgctcttgt ccttggcttt cttcctctta gctcgctctc ggctgctgga gcggtcactg
     4321 gaggaccgcc ggctcttggc cttggaccgc tgcctcctgg ggcgctcctc acccactgat
     4381 ggtgacgctg acctcgagct cctgtccctg gattcagagc gcgtcccaga gcgcgtccgt
     4441 ccgtgctccc tggacaccct cttcctcttg gccttcttcc tgctgcggga gctggatgtg
     4501 gagcgggacc gggaaggggg cctgagctcc acgactctgt gggtggcagc ctgcagcacg
     4561 tccgggctgg gtggggcttc cggctccacg acagtcacac aggtcaccgt ccgctcctca
     4621 gagccaaaga aggtgctgca agacgcccca tcctcctcat cccagggctc tgggggggat
     4681 ggcctccgca gccgggcagc caggccctgg ctctctgtgg gctcctctcg ctccgtgtca
     4741 gaggccccct cggcagccac tgcaccccgg agcttggaca tggcggagga cggtccgaga
     4801 gggggttcgg gctctgtacc aaagatgctc tccaccgtgt aggaggtgac gcagcgcaca
     4861 gcctgcaccg tctgggcctt cgggctgttg atggagatgg ttcgtgtgat ctcagagggc
     4921 aggaggccgg ggccagacct ctcggggctg ctgccagggg cgctggagtc ggagccggtg
     4981 gggtcggatg ggtcgtagac ctcgctccgc agctgcttcg tcttcttgat ggagaagagg
     5041 ggtgaggggt tctccttcct ctgctcctta tcgtccacag gccggaacgt gttacagaag
     5101 ccagggctgg agagctggct ggaatgtgcc atgtttccag gaatattgat ccggaagctc
     5161 tcaaaggtcg acccgacccc tttccctctt gatggtccca gtggggccaa actgccatgg
     5221 gagctggggg ctgggggccg ggagctgccc gtgtgcaccc cgggctcccg gggcaccctg
     5281 ctgctggcct cgctctctgc ccgtgtgcct cgccctggct gcccggtgcc ctccctgcca
     5341 gtcagtcggc tgatgcaggc actggggatc tcaatgctct gcccgtgggc cggggccgca
     5401 tcccgtctgc caccaccgtc atctctcctg atctttggta tcctgggtag ctcagagatg
     5461 tctgttcttc tgggcgctgg cttcagggga ggccccggca ccacactgcg acatgggggc
     5521 ttagaatctc tgcttgagat cttagacgcg gcagaggcaa ggggcaaggt gtgcttgttg
     5581 gtgccgttga accaggggct gtggctctgt ctgaagccag gcacactccc atttgacaag
     5641 ttccgagcct gaaccgcccc aggggctgct ggcaagtcca gcctcacagg cgcccccgcg
     5701 gtgcgggcgg gggtgcggga cctgccttga cagctgagcc ctgtgctctg tggcctgttt
     5761 cctgacgggc cttgggccgg gctcccggag ggcagtgctc ggggctgcag gcagcccttg
     5821 aatccttctt cccctctgga cagactgcct gagtttcgct gaaaagaaac tggagccgcc
     5881 ctcttggcgc tgagggagcc gtcgcggtgg atgatgacat cactgctgcc cagcatcagg
     5941 aggctctggc ccgacaggat gctgcccagc aggtcaggca ctggctcctc ctccaagtct
     6001 ggctctggca ccgcggcagg gaggcgcctc ctggaaagcc ccacggagac gggcctggcc
     6061 acgggctggt gggactgcag ggctgaccgg gacagagccc gtctcttggc actcagaggg
     6121 gaaagagggt ttgcagaaag ctcttcactg ctgtcgaagg gatccagctc atagggatct
     6181 ccaaacagag acagagaggc agctccaata tccgctctca gcagccccaa agagggctcc
     6241 actggcttca acactgacgg gatgcagctg ctgtgaacag gcctgcgcag gcccagcgtc
     6301 cgcgcgattc gagagcgagt ggtggcttca ctctttacct tcttccctct tctcttcttc
     6361 actcgatgtt ggcgtttctt agatcttgtc gctgagctct tactttttgc ggatggtccg
     6421 gacggggttt tctttcttcc cggcactttc ttccgtcttc ttgtcttcct cttccgtcgg
     6481 gcgggagtgc gcggcgtcag ggggcgctga tacacggcag tgctcaggcc agtcgccaca
     6541 gcctcgatgg cttcatccag cagggaagac ccgaggcgcc ctggtgtgtg ctggaccctc
     6601 ctggccgtgg agatccggtt ccggttcacg gttgctctca ctctctcact ctgccgtgtc
     6661 ctggctatcg cccgggtcct acctgctcga ggccgaagcc tgctggtggt gggcaccaca
     6721 tcagccaaga gcagggagac ctcctcctca ctcacgggac ccgcatcagc ggcaaggaca
     6781 acaccaggcg cagcacattc cgggcagaac cactcgt
//
