LOCUS       sequence-491880-        6576 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for HECW2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6576)
  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  JOURNAL   Unpublished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 6576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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'-HECW2
FEATURES             Location/Qualifiers
     source          1..6576
                     /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   3850..3979
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      4054..4509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4141..4160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4351..4372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4526..4545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4645..4667
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4705..4723)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4791..4895
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4896..4964,4965..5756)
                     /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(5114..5133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5927..6515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6416..6435
                     /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 ggatctgcta gttcagcccg ggagcacctg ctttttgtga ggcgtcgaaa
      781 tccccagatg cggtacacat tgagcccaga gaacctccag agccttgccg cccagagctc
      841 catgccagag aacatgaccc tgcagcgggc caacagcgac accgacctgg tgacttctga
      901 gagccgctcc agcttaactg ccagcatgta cgagtacacg ctggggcaag cccagaacct
      961 cattatcttc tgggacatta aagaggaggt ggaccccagt gattggattg gactttatca
     1021 tatagatgag aattctccag ccaacttctg ggattctaag aacaggggtg tgactggaac
     1081 acaaaaaggg caaattgtat ggagaattga gcctgggccc tatttcatgg aaccggagat
     1141 aaaaatctgt tttaaatatt accacggcat tagtggagcc ctgcgagcca cgaccccctg
     1201 catcaccgtg aagaacccag ctgtgatgat gggggcagaa ggcatggagg gaggtgcttc
     1261 aggaaacctg cattctcgaa aacttgttag ctttacattg tcagatctta gggcagttgg
     1321 gctaaagaaa gggatgttct tcaatcctga cccttatctt aagatgtcaa ttcagccagg
     1381 aaagaagagc agtttcccca cctgtgccca ccacgggcag gagagacggt ctactatcat
     1441 cagtaacacc accaatccaa tttggcaccg agagaaatat tccttttttg cacttcttac
     1501 tgatgtctta gaaattgaaa ttaaagacaa atttgccaag agccgtccca tcatcaagcg
     1561 ttttctgggg aaactaacca ttccagtcca gaggctgctg gagcgacaag ccatcggtga
     1621 tcaaatgctc agctacaacc ttggcagaag gctcccagct gaccacgtga gtgggtacct
     1681 ccagtttaaa gtggaggtta cgtcttctgt tcatgaagat gcctctccag aagctgttgg
     1741 cacaatactt ggagtcaatt ctgtgaatgg agacttaggt agcccttccg atgacgagga
     1801 catgccaggg agccatcacg acagccaggt gtgctctaat gggccagttt ctgaggacag
     1861 tgctgccgat ggaaccccca agcattcatt caggactagc tccactctgg aaatagacac
     1921 agaggaatta acctctacgt cttcaaggac ctcacctccc agaggacgtc aggattcact
     1981 caatgattac ttagatgcta tcgaacacaa tggccactcc aggccaggga cagcgacctg
     2041 ctctgagagg tccatgggtg cctctccgaa actgaggagt agttttccca ctgacacaag
     2101 actcaatgcc atgcttcata ttgattcaga tgaagaagat cacgagttcc agcaagacct
     2161 gggttaccca tcttccttgg aggaggaggg aggcctgatc atgtttagca gggcatccag
     2221 agctgatgat ggaagcctga catctcagac aaagctggag gacaaccctg ttgagaatga
     2281 ggaagcctcc acacacgaag ctgcttcctt tgaggataag ccagaaaatc ttccagagct
     2341 tgcagagagc tccttacctg caggcccagc cccagaggaa ggtgaaggcg gcccagagcc
     2401 tcaacccagt gctgaccagg gcagtgctga actgtgtggc tctcaagagg tagatcagcc
     2461 cacaagtggc gcagacacag ggacttccga tgcatcagga ggaagccgaa gagccgtcag
     2521 tgagaccgag tctctcgatc agggctctga gccttcccag gtgtcctctg aaacagaacc
     2581 cagtgatcct gccaggacag agagtgtgag cgaagccagc accaggcctg agggagagag
     2641 tgacctggaa tgcgctgaca gctcctgcaa tgagagtgtg accacgcagc tgtcctctgt
     2701 ggacacacgg tgctcctctc ttgagagcgc acggtttcct gaaaccccag ccttttcttc
     2761 tcaggaggag gaagacggag cctgtgcagc agagcccacc agcagtggcc ctgccgaagg
     2821 gtcgcaggaa tccgtgtgca ctgctggttc tttacctgtg gtacaggtgc ccagtgggga
     2881 ggatgaaggg ccaggggcag aatcggccac tgtacctgac caggaggagc tgggggaggt
     2941 ctggcagcgg agggggagcc tggagggagc tgcagctgct gccgagagcc caccgcaaga
     3001 agaaggcagt gctggggagg cccaaggcac ctgtgaaggg gcaactgccc aggaggaggg
     3061 cgctactgga ggttctcaag ccaacggcca ccagccactg cgatcactac cttcagtgcg
     3121 caaggatgtt agccggtacc agagggtgga cgaggctctc ccaccaaact gggaggcacg
     3181 cattgacagc cacggcagga tcttctacgt ggatcacgta aacagaacca cgacgtggca
     3241 gcgaccgaca gctcccccag ccccgcaggt gctgcagaga tctaactcca tacagcaaat
     3301 ggagcagctg aaccggcgat atcagagtat ccgcagaacc atgaccaatg agaggcctga
     3361 ggaaaatacc aatgctattg atggggcagg ggaggaagct gactttcacc aagccagtgc
     3421 agatttccga cgggagaaca tcctgcctca ctctacctcc agatccagga tcacgttgct
     3481 gttacagtct ccccccgtga agttcctcat cagcccagag ttcttcaccg tgctgcattc
     3541 taaccctagt gcctaccgca tgtttacaaa caacacgtgt ttgaagcaca tgatcaccaa
     3601 agtccggagg gacacccacc actttgaacg ctaccagcat aaccgcgacc ttgtgggatt
     3661 cctcaacatg ttcgcgaaca aacagctaga gctgccgtgg ggatgggaaa tgaaacatga
     3721 tcaccaggta agtatcaagg ttacaagaca ggtttaagga gaccaataag ctattattcg
     3781 tataggatac tttatacgaa gttattaagt tatggagacc ggtgtggcac gtgcggaccg
     3841 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3901 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga
     3961 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4021 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     4081 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     4141 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4201 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4261 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4321 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4381 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     4441 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4501 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4561 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4621 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4681 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4741 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4801 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4861 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4921 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4981 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5041 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5101 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5161 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5221 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5281 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5341 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5401 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5461 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5521 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5581 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5641 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5701 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5761 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5821 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5881 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5941 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6001 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6061 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6121 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6181 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6241 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6301 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6361 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6421 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6481 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6541 ggttcctggc cttttgctgg ccttttgctc acatgt
//
