LOCUS       sequence-488703-        6834 bp    DNA     circular SYN 08-OCT-2025
DEFINITION  5' AAVLINK plasmid for RERE expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6834)
  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  JOURNAL   Unpublished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 6834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 8, 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            "Unpublished"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'-RERE
FEATURES             Location/Qualifiers
     source          1..6834
                     /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        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     RBS             1722..1730
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     repeat_region   4097..4237
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4097..4226
                     /label=AAV2 ITR
     rep_origin      4312..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4399..4418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4609..4630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4784..4803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4903..4925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4963..4981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5154..5222,5223..6014)
                     /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(5372..5391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctac agcggacaaa gacaaagaca aagacaaaga
      781 gaaggaccgg gaccgagacc gggaccgaga gagagagaaa agagacaaag caagagagag
      841 tgagaattca aggccacgcc ggagctgtac cttggaagga ggagccaaaa attatgctga
      901 gagtgatcac agtgaagacg aggacaatga caacaatagt gccaccgcag aggagtccac
      961 gaagaagaat aagaagaaac caccgaaaaa aaagtctcgt tatgaaagga cagataccgg
     1021 tgagataaca tcctacatca ctgaagatga tgtggtctac agaccaggag actgtgtgta
     1081 tatcgagagt cggaggccaa acacaccgta tttcatctgt agcattcaag acttcaaact
     1141 ggtccacaac tcccaggcct gttgcagatc tccaactcct gctttgtgtg accccccagc
     1201 atgctctctg ccggtggcat cacagccacc acagcatctt tctgaagccg ggagagggcc
     1261 tgtagggagt aagagggacc atctcctcat gaacgtcaaa tggtactacc gtcaatctga
     1321 ggttccagat tctgtgtatc agcatttggt tcaggatcga cataatgaaa atgactctgg
     1381 aagagaactt gtcattacag acccagttat caagaaccga gagctcttca tttctgatta
     1441 cgttgacact taccatgctg ctgcccttag agggaagtgt aacatctccc atttttctga
     1501 catatttgct gctagagagt ttaaagcccg agtggattca tttttctaca tattaggata
     1561 taaccctgag acaaggaggc tgaacagtac ccagggggag attcgtgtcg gtcctagtca
     1621 tcaggccaaa cttccagatc tgcaaccatt tccttctcca gatggtgata cagtgaccca
     1681 acatgaggaa ctggtctgga tgcctggagt taacgactgt gacctcctta tgtacttgag
     1741 ggcagcaagg agcatggcgg catttgcagg aatgtgtgat ggaggctcta cagaggacgg
     1801 ctgtgtcgca gcctctcggg atgacaccac tctgaatgca ctgaacacac tgcatgaaag
     1861 cggttacgat gctggcaaag ccctgcagcg cctggtgaag aagcctgtgc ccaagctcat
     1921 cgagaagtgc tggaccgagg acgaagtgaa acgcttcgtt aagggactca ggcagtacgg
     1981 gaagaacttc ttcagaatta gaaaggagct gcttcccaat aaggaaacag gggagctgat
     2041 caccttctat tactattgga agaagacccc cgaagcagcc agctcccgag cccatcgtag
     2101 gcaccgcagg caggccgtgt tcaggaggat taagactcgc accgcgtcca cacccgtcaa
     2161 cacaccctcc agacccccgt ccagtgaatt cttggaccta agttcagcca gtgaagatga
     2221 cttcgacagt gaggacagtg agcaggagct gaaggggtac gcctgccgcc actgcttcac
     2281 caccacctcc aaagattggc accacggagg ccgggagaac atcctgcttt gcaccgactg
     2341 tcgcatccac ttcaagaaat acggtgagct cccgcccatt gagaagcccg tggacccgcc
     2401 accgtttatg ttcaaacccg tcaaggaaga ggatgatggg ctcagtggga agcatagcat
     2461 gaggacacgg cggagtcggg gctcgatgtc gacactacgc agtggtcgga agaagcagcc
     2521 agccagccct gatggtcgca cctcacccat caatgaagac atccgctcca gcggccggaa
     2581 ctcccccagc gctgccagta cctccagcaa tgacagtaaa gcagagacag tgaagaagtc
     2641 ggccaagaag gtgaaggagg aagcctcttc ccctcttaag agtaacaaac gccagcggga
     2701 gaaggtggcc tctgatacgg aggaggctga caggaccagc tccaagaaga caaaaacgca
     2761 ggagatcagc aggcccaact cgccatctga aggtgaggga gagagttcag acagtcgcag
     2821 cgtcaacgat gagggtagca gtgaccccaa agacatcgac caggacaatc gcagcacgtc
     2881 cccgagcatc cccagccccc aggacaatga gagtgactcg gactcgtcag cccagcagca
     2941 gatgctgcag gcccagcccc cagccttgca ggctcccact ggggtcaccc cagctccctc
     3001 ctcagctcct ccagggaccc ctcagctgcc cacgccaggg cccacgccct ctgccactgc
     3061 agttccccca cagggctccc ccacggcctc ccaggcccct aaccagccac aggctcccac
     3121 agcgcctgtt ccccacaccc acatccaaca ggcaccggcc ttgcaccccc agcggccgcc
     3181 ctcaccgcat cccccgccgc atccctcgcc acatcccccg ctgcagcctc tgactgggtc
     3241 ggcgggccag ccttctgcac cctctcatgc ccagccccca ctgcacggtc agggcccacc
     3301 cggccctcac agcctgcagg ctgggcccct gctgcagcac ccaggccccc cacagccctt
     3361 tggcctccct ccccaggcct cccaaggcca ggcccctctg gggacctccc cagcagcagc
     3421 gtaccctcac acctccctgc agctgccagc ctctcagtca gcgctgcagt cccaacagcc
     3481 tccacgggag cagcccctgc caccagcgcc cttggccatg ccccacatca agcccccgcc
     3541 taccactccc atcccccagc tgccggcgcc acaggcccac aagcaccctc cccacctctc
     3601 ggggccctca cccttctcca tgaatgccaa cctgcctccc cctccagccc tgaagcccct
     3661 gagctccctg tccacacatc accccccgtc ggctcacccc ccacccctgc aactcatgcc
     3721 tcagagccag ccattgccct cctcgcccgc ccagcccccc gggctgaccc agagccagaa
     3781 cctgcccccg ccccctgcct cccacccccc tacaggcctc caccaggtgg ccccccaacc
     3841 cccgtttgct cagcacccct ttgtccctgg aggccctcct cccatcaccc ctccgacctg
     3901 cccctccacc tctaccccac cggcgggacc tggcacctcg gcccagccac cctgctctgg
     3961 tgcggcggct tcaggtaagt atcaaggtta caagacaggt ttaaggagac caataagcta
     4021 ttattcgtat aggatacttt atacgaagtt attaagttat ggagaccggt gtggcacgtg
     4081 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4141 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4201 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4261 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4381 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4441 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4501 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4621 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     4681 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4741 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4801 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4861 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4921 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
