LOCUS       sequence-487598-        5659 bp    DNA     circular SYN 01-OCT-2025
DEFINITION  5' AAVLINK plasmid for BRD4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5659)
  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  JOURNAL   Unpublished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 5659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 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            "Unpublished"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' BRD4
FEATURES             Location/Qualifiers
     source          1..5659
                     /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"
     rep_origin      3137..3592
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3224..3243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3434..3455
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3609..3628)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3728..3750
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3788..3806)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3874..3978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3979..4047,4048..4839)
                     /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(4197..4216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5010..5598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5499..5518
                     /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 ggatcttctg cggagagcgg ccctgggacg agattgagaa atctgccagt
      781 aatgggggat ggactagaaa cttcccaaat gtctacaaca caggcccagg cccaacccca
      841 gccagccaac gcagccagca ccaacccccc gcccccagag acctccaacc ctaacaagcc
      901 caagaggcag accaaccaac tgcaatacct gctcagagtg gtgctcaaga cactatggaa
      961 acaccagttt gcatggcctt tccagcagcc tgtggatgcc gtcaagctga acctccctga
     1021 ttactataag atcattaaaa cgcctatgga tatgggaaca ataaagaagc gcttggaaaa
     1081 caactattac tggaatgctc aggaatgtat ccaggacttc aacactatgt ttacaaattg
     1141 ttacatctac aacaagcctg gagatgacat agtcttaatg gcagaagctc tggaaaagct
     1201 cttcttgcaa aaaataaatg agctacccac agaagaaacc gagatcatga tagtccaggc
     1261 aaaaggaaga ggacgtggga ggaaagaaac agggacagca aaacctggcg tttccacggt
     1321 accaaacaca actcaagcat cgactcctcc gcagacccag acccctcagc cgaatcctcc
     1381 tcctgtgcag gccacgcctc accccttccc tgccgtcacc ccggacctca tcgtccagac
     1441 ccctgtcatg acagtggtgc ctccccagcc actgcagacg cccccgccag tgccccccca
     1501 gccacaaccc ccacccgctc cagctcccca gcccgtacag agccacccac ccatcatcgc
     1561 ggccacccca cagcctgtga agacaaagaa gggagtgaag aggaaagcag acaccaccac
     1621 ccccaccacc attgacccca ttcacgagcc accctcgctg cccccggagc ccaagaccac
     1681 caagctgggc cagcggcggg agagcagccg gcctgtgaaa cctccaaaga aggacgtgcc
     1741 cgactctcag cagcacccag caccagagaa gagcagcaag gtctcggagc agctcaagtg
     1801 ctgcagcggc atcctcaagg agatgtttgc caagaagcac gccgcctacg cctggccctt
     1861 ctacaagcct gtggacgtgg aggcactggg cctacacgac tactgtgaca tcatcaagca
     1921 ccccatggac atgagcacaa tcaagtctaa actggaggcc cgtgagtacc gtgatgctca
     1981 ggagtttggt gctgacgtcc gattgatgtt ctccaactgc tataagtaca accctcctga
     2041 ccatgaggtg gtggccatgg cccgcaagct ccaggatgtg ttcgaaatgc gctttgccaa
     2101 gatgccggac gagcctgagg agccagtggt ggccgtgtcc tccccggcag tgccccctcc
     2161 caccaaggtt gtggccccgc cctcatccag cgacagcagc agcgatagct cctcggacag
     2221 tgacagttcg actgatgact ctgaggagga gcgagcccag cggctggctg agctccagga
     2281 gcagctcaaa gccgtgcacg agcagcttgc agccctctct cagccccagc agaacaaacc
     2341 aaagaaaaag gagaaagaca agaaggaaaa gaaaaaagaa aagcacaaaa ggaaagagga
     2401 agtggaagag aataaaaaaa gcaaagccaa ggaacctcct cctaaaaaga cgaagaaaaa
     2461 taatagcagc aacagcaatg tgagcaagaa ggagccagcg cccatgaaga gcaagccccc
     2521 tcccacgtat gagtcggagg aagaggacaa gtgcaagcct atgtcctatg aggagaagcg
     2581 gcagctcagc ttggacatca acaagctccc cggcgagaag ctgggccgcg tggtgcacat
     2641 catccagtca cgggagccct ccctgaagaa ttccaacccc gacgagattg aaatcgactt
     2701 tgagaccctg aagccgtcca cactgcgtga gctggagcgc tatgtcacct cctgtttgcg
     2761 gaagaaaagg aaacctcaag ctgagaaagt tgatgtgatt gccggctcct ccaagatgaa
     2821 ggtaagtatc aaggttacaa gacaggttta aggagaccaa taagctatta ttcgtatagg
     2881 atactttata cgaagttatt aagttatgga gaccggtgtg gcacgtgcgg accgagcggc
     2941 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3001 aggccgggcg accaaaggtc gcccggcctc agtgagcgag cgagcgcgca gctgcctgca
     3061 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3121 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3181 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3241 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3301 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3361 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3421 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3481 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3541 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3601 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3661 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3721 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3781 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3841 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3901 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3961 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4021 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4081 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4141 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4201 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4261 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4321 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4381 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4441 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4501 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4561 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4621 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4681 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4741 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4801 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4861 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4921 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5641 tggccttttg ctcacatgt
//
