LOCUS       sequence-491793-        6034 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for ERBIN expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  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  JOURNAL   Unpublished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 6034)
  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            "Unpublished"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'-ERBIN
FEATURES             Location/Qualifiers
     source          1..6034
                     /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      3512..3967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3599..3618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3809..3830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3984..4003)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4103..4125
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4163..4181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4249..4353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4354..4422,4423..5214)
                     /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(4572..4591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5385..5973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5874..5893
                     /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 ggatctacta caaaacgaag tttgtttgtg cggttggtac catgtcgctg
      781 tctacgaggg gaagaggaga ctgtcactac tcttgattat tctcattgca gcttagaaca
      841 agttccgaaa gagattttta cttttgaaaa aaccttggag gaactctatt tagatgctaa
      901 tcagattgaa gagcttccaa agcaactttt taactgtcag tctttacaca aactgagttt
      961 gccagacaat gatttaacaa cgttaccagc atccattgca aaccttatta atctcaggga
     1021 actggatgtc agcaagaatg gaatacagga gtttccagaa aatataaaaa attgtaaagt
     1081 tttgacaatt gtggaggcca gtgtaaaccc tatttccaag ctccctgatg gattttctca
     1141 gctgttaaac ctaacccagt tgtatctgaa tgatgctttt cttgagttct tgccagcaaa
     1201 ttttggcaga ttaactaaac tccaaatatt agagcttaga gaaaaccagt taaaaatgtt
     1261 gcctaaaact atgaatagac tgacccagct ggaaagattg gatttgggaa gtaacgaatt
     1321 cacggaagtg cctgaagtac ttgagcaact aagtggattg aaagagtttt ggatggatgc
     1381 taatagactg acttttattc cagggtttat tggtagtttg aaacagctca catatttgga
     1441 tgtttctaaa aataatattg aaatggttga agaaggaatt tcaacatgtg aaaaccttca
     1501 agacctccta ttatcaagca attcacttca gcagcttcct gagactattg gtttgttgaa
     1561 gaatataaca acgcttaaaa tagatgaaaa ccagttaatg tatctgccag actctatagg
     1621 agggttaata tcagtagaag aactggattg tagtttcaat gaagttgaag ctttgccttc
     1681 atctattggg cagcttacta acttaagaac ttttgctgct gatcataatt acttacagca
     1741 gttgccccca gagattggaa gctggaaaaa tataactgtg ctgtttctcc attccaataa
     1801 acttgagaca cttccagagg aaatgggtga tatgcaaaaa ttaaaagtca ttaatttaag
     1861 tgataataga ttaaagaatt taccctttag ctttacaaag ctacagcaat tgacagctat
     1921 gtggctctca gataatcagt ccaaacccct gatacctctt caaaaagaaa ctgattcaga
     1981 gacccagaaa atggtgctta ccaactacat gttccctcaa cagccaagga ctgaggatgt
     2041 tatgtttata tcagataatg aaagttttaa cccttcattg tgggaggaac agaggaaaca
     2101 gcgggctcaa gttgcatttg aatgtgatga agacaaagat gaaagggagg cacctcccag
     2161 ggagggaaat ttaaaaagat atccaacacc atacccagat gagcttaaga atatggtcaa
     2221 aactgttcaa accattgtac atagattaaa agatgaagag accaatgaag actcaggaag
     2281 agatttgaaa ccacatgaag atcaacaaga tataaataaa gatgtgggtg tgaagacctc
     2341 agaaagtact actacagtaa aaagcaaagt tgatgaaaga gaaaaatata tgataggaaa
     2401 ctctgtacag aagatcagtg aacctgaagc tgagattagt cctgggagtt taccagtgac
     2461 tgcaaatatg aaagcctctg agaacttgaa gcatattgtt aaccatgatg atgtttttga
     2521 ggaatctgaa gaactttctt ctgatgaaga gatgaaaatg gcggagatgc gaccaccatt
     2581 aattgaaacc tctattaacc agccaaaagt cgtagcactt agtaataaca aaaaagatga
     2641 tacaaaggaa acagattctt tatcagatga agttacacac aatagcaatc agaataacag
     2701 caattgttct tctccatctc ggatgtctga ttcagtttct cttaatactg atagtagtca
     2761 agacacctca ctctgctctc cagtgaaaca aactcatatt gatattaatt ccaaaatcag
     2821 gcaagaagat gaaaatttta acagcctttt acaaaatgga gatattttaa acagttcaac
     2881 agaggaaaag ttcaaagctc atgataaaaa agattttaac ttacctgaat atgatttgaa
     2941 tgttgaagag cgattagttc taattgagaa aagtgttgac tcaacagcca cagctgatga
     3001 cactcacaaa ttagatcata tcaatatgaa tcttaataaa cttataacta atgatacatt
     3061 tcaaccagag atcatggaaa gatcaaaaac acaggatatt gtgcttggaa caagcttttt
     3121 aagcattaat tctaaagagg aaactgagca cttggaaaat ggaaacaagt atcctaattt
     3181 ggaatccgta aataaggtaa gtatcaaggt tacaagacag gtttaaggag accaataagc
     3241 tattattcgt ataggatact ttatacgaag ttattaagtt atggagaccg gtgtggcacg
     3301 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3361 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg gcctcagtga gcgagcgagc
     3421 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3481 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3541 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3601 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3661 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3721 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3781 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3841 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3901 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3961 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4021 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4081 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4141 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4201 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4261 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4321 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4381 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4441 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4501 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4561 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4621 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4681 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4741 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4801 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4861 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4921 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4981 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5041 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5101 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5161 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5221 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5281 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6001 ttcctggcct tttgctggcc ttttgctcac atgt
//
