LOCUS       sequence-491864-        6105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for CECR2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6105)
  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  JOURNAL   Unpublished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 6105)
  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            "Unpublished"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-HA-5'-CECR2
FEATURES             Location/Qualifiers
     source          1..6105
                     /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   3368..3508
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3368..3497
                     /label=AAV2 ITR
     rep_origin      3583..4038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3670..3689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3880..3901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4055..4074)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4174..4196
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4234..4252)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4320..4424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4425..4493,4494..5285)
                     /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(4643..4662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5456..6044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5945..5964
                     /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 ggatcttgcc cagaggaggg cggcgcggcc gggctgggcg agctccgctc
      781 ctggtgggag gtcccggcca tcgcgcactt ctgctcgctc tttcgcaccg cgttccgcct
      841 gcccgacttc gagatcgagg agttagaagc cgctcttcac agagatgacg tggagtttat
      901 cagtgacctg attgcctgcc tgcttcaggg ctgctatcaa cgaagagata tcacgcctca
      961 gacattccac agctacctag aggacatcat caactaccgc tgggagctcg aagaagggaa
     1021 gcccaaccct ctgagggaag ccagtttcca ggacctgcct cttcgcacac gggtggagat
     1081 cctgcaccga ctctgtgatt accggctgga tgcagacgat gtcttcgatc ttctaaaggg
     1141 cctggatgca gacagtctcc gtgtggagcc attgggtgaa gacaattctg gggcactata
     1201 ttggtatttc tatggaacac gaatgtacaa agaggacccg gtgcaaggaa aatccaatgg
     1261 agaactctct ttgagcaggg aaagtgaagg acaaaaaaat gtctcaagta ttcctggaaa
     1321 aacgggaaaa agaagaggaa gacccccaaa acggaagaaa ctgcaggagg agattctgtt
     1381 gagtgaaaag caggaagaaa attccttggc atccgagcca cagacaagac atgggtccca
     1441 agggccaggc caaggtactt ggtggctcct gtgccagaca gaagaggaat ggagacaggt
     1501 caccgagagt tttcgcgaga ggacctccct tcgagaacgg cagctctaca agctcctcag
     1561 tgaggacttc ctgcctgaga tctgcaacat gatcgcccag aagggaaaac gtccacagcg
     1621 cacaaaggca gagttgcatc ctaggtggat gtctgaccac ctgtccatca aacccgtcaa
     1681 gcaagaggag actcctgtgc tgaccagaat agaaaaacaa aagcgcaaag aggaggaaga
     1741 agagcgtcag attcttctag cagtgcagaa gaaggagcag gagcagatgc taaaggaaga
     1801 gaggaaacgc gagttggagg agaaggtcaa ggcagtggaa gatcgagcga agaggagaaa
     1861 gctcagggaa gaaagggcat ggctgctggc tcaaggaaag gagctccctc cagaactttc
     1921 ccatctggac cccaattccc ccatgagaga ggaaaaaaag actaaagacc tctttgagtt
     1981 ggatgatgat ttcactgcta tgtataaagt tctagacgtg gtaaaggctc acaaggattc
     2041 ctggcccttc ttggaacctg tggatgaatc ttatgcccct aactattatc agattattaa
     2101 ggcccccatg gatatttcca gcatggagaa gaaactgaat ggaggtttat actgtaccaa
     2161 ggaggaattt gtaaatgaca tgaagaccat gttcaggaat tgtcgaaagt ataatgggga
     2221 aagtagtgag tataccaaga tgtctgataa tttagagagg tgtttccatc gggcaatgat
     2281 gaaacatttt cctggagaag atggagacac agatgaagaa ttttggattc gagaggatga
     2341 aaagcgggag aaaagacgga gtcgggctgg gcgaagtggt gggagccatg tttggacccg
     2401 ctccagggac ccagaagggt ccagcaggaa acagcagccc atggagaatg gaggaaagtc
     2461 gttgcccccc acacgccgag cgccctcttc tggggacgat cagagcagca gctccacaca
     2521 gcccccgcgg gaggtgggca cttccaatgg ccgaggtttt tctcatcccc tgcattgtgg
     2581 tgggacaccc agccaggcac cctttttaaa ccagatgagg ccagcagtac caggaacatt
     2641 tggccctctg cgaggatcag atcctgccac cttgtatggc tcctctggag tcccggagcc
     2701 acaccacggg gagcctgtgc agcagcgtca gcctttcacc atgcagcctc cagttggaat
     2761 taacagcctc cgaggaccca ggctaggcac accagaggag aagcaaatgt gcggggggct
     2821 gacacacctt tctaacatgg gcccacaccc tggatccttg cagcttgggc agataagtgg
     2881 cccaagtcag gatggaagca tgtatgctcc agctcagttc cagccaggat tcattcctcc
     2941 ccggcatggg ggggctccag cccggccacc agactttcct gaaagctcag aaattcctcc
     3001 cagccatatg tatcgatcgt acaagtacct gaatcgagta cactctgccg tctggaatgg
     3061 gaaccatggt gctacgaacc aaggaccctt gggcccagat gagaagcccc acctggggcc
     3121 aggaccctct caccagcctc gcactctcgg tcacgtgatg gattcccgag tcatgagacc
     3181 acctgtcccc cccaaccagt ggactgaaca atcaggcttc ctacctcatg gagttccttc
     3241 ctcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaataagct attattcgta
     3301 taggatactt tatacgaagt tattaagtta tggagaccgg tgtggcacgt gcggaccgag
     3361 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3421 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3481 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     3541 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3601 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     3661 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3721 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3781 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3841 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3901 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     3961 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4021 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4081 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4141 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4201 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4261 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4441 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4501 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4561 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4621 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4681 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4741 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4801 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4861 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4921 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4981 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5041 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5101 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5161 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5221 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5281 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5341 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5581 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6061 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
