LOCUS       sequence-491877-        6163 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for DIP2C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6163)
  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  JOURNAL   Unpublished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 6163)
  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            "Unpublished"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'-DIP2C
FEATURES             Location/Qualifiers
     source          1..6163
                     /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      3641..4096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3728..3747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3938..3959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4113..4132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4232..4254
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4378..4482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4483..4551,4552..5343)
                     /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(4701..4720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5514..6102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6003..6022
                     /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 ggatctgcgg accgcagcct ggagggcatg gcgctgcccc tggaggtgcg
      781 ggcgcgcctg gccgagctgg agctggagct gtcggaaggt gacatcacac aaaaaggata
      841 tgaaaagaag aggtcaaagt taattggagc ctaccttccg cagcctccga gggtggacca
      901 agctttgccg caagaacgcc gggctcctgt cactccttcc tccgcctctc gctaccaccg
      961 ccgacggtct tcagggtcac gagatgagcg ctatcggtca gacgtccaca cggaagctgt
     1021 ccaggcggct ctggccaaac acaaagagcg gaagatggca gtgcctatgc cttccaaacg
     1081 caggtccctg gtcgtgcaga cctcgatgga cgcctacacc cctccagata cctcttctgg
     1141 ctcagaagat gaaggctcag tgcaggggga ctcccagggc acccccacct ccagccaggg
     1201 cagcatcaat atggagcact ggatcagcca ggccatccac ggctccacca cgtccaccac
     1261 ctcctcgtcc tctacgcaga gcgggggcag cggggctgcc cacaggctgg cggacgtcat
     1321 ggctcagacc cacatagaaa atcattctgc acctcctgac gtaaccacgt acacctcaga
     1381 gcactcgata caggtggaga gaccgcaggg ttccacgggg tcccggacag cgcccaagta
     1441 cggcaacgcc gagctcatgg agaccgggga tggagtacca gtaagtagcc gggtgtcagc
     1501 aaaaatccag cagcttgtca ataccctcaa acgaccgaaa cgaccacctt tacgagaatt
     1561 ctttgtcgat gactttgaag aattattaga agttcaacaa ccggatccga accaaccaaa
     1621 gccggagggg gcccagatgc tggccatgcg cggagagcag ctgggcgtgg tcacgaactg
     1681 gccgccgtcg ctggaggccg cactgcagag gtggggcacc atctcgccca aggcgccctg
     1741 cctgaccacc atggacacca acgggaagcc cctctacatc ctcacttacg gcaagctgtg
     1801 gacaagaagt atgaaggtcg cttacagcat tctacacaaa ttaggcacaa agcaggaacc
     1861 catggtccgg cctggagata gggtggcact ggtgttcccc aacaatgatc cggctgcctt
     1921 catggcggct ttctacggct gcctgctggc cgaggtggtc cccgtgccca tcgaggtgcc
     1981 actcaccagg aaggacgcag ggagccagca gataggtttc ttgcttggaa gctgtggagt
     2041 tactgtagcc ttgactagtg acgcctgcca taaaggactt ccaaaaagcc caacgggaga
     2101 gatcccacag tttaaaggtt ggccaaagct gctgtggttt gtcacagagt ctaaacatct
     2161 ctccaaaccg ccccgagact ggttcccaca cattaaagat gccaataacg acactgcgta
     2221 tattgagtac aagacgtgta aggatggcag tgtgctgggt gtgacggtga cgaggactgc
     2281 gctgctgaca cactgccagg ccctgacgca ggcgtgtggc tacacggaag ctgaaaccat
     2341 tgtgaatgtg ctggatttca agaaggacgt cgggctctgg catggcatcc tgacaagcgt
     2401 catgaacatg atgcatgtga tcagcatccc gtactcgctg atgaaggtga accctctctc
     2461 ctggatccag aaggtctgcc agtacaaagc aaaagtggcg tgtgtgaaat cgagggatat
     2521 gcattgggca ttagtagcac acagagatca gagagacatc aacctctcct ctctgcgaat
     2581 gctgatagtg gcggacggcg cgaacccctg gtctatttct tcttgcgatg catttctcaa
     2641 tgtcttccaa agtaaaggcc ttcgacagga ggtcatctgt ccttgtgcca gctcgccaga
     2701 ggccctcact gtggccatcc ggaggcccac ggatgacagt aaccagcccc cgggccgggg
     2761 tgtcctctcc atgcatggac tgacctatgg ggtcattcgt gtggactcgg aagagaagct
     2821 gtccgtgctc accgtgcagg atgtcggcct cgtgatgcct ggagccatca tgtgttcagt
     2881 gaagccagac ggggttcctc agctgtgcag aacggatgag atcggggagc tgtgtgtgtg
     2941 tgcagttgcg acgggcacgt cctactatgg cctctctggc atgaccaaga acacctttga
     3001 ggtgtttccc atgacaagct ccggggctcc gatcagtgaa tacccattca taaggacagg
     3061 cttgctgggg ttcgtgggtc ccggaggcct cgtcttcgtg gtgggcaaga tggatggcct
     3121 catggtggtc agcgggcgca ggcacaacgc cgacgacatc gtggccactg cgctggccgt
     3181 agaacccatg aagtttgtct accggggaag gatagccgtg ttctcggtga ccgtgctgca
     3241 cgacgagagg atcgtgatcg tggctgagca gaggcctgac tccacggaag aggacagttt
     3301 ccagtggatg agccgtgtgc tgcaggtaag tatcaaggtt acaagacagg tttaaggaga
     3361 ccaataagct attattcgta taggatactt tatacgaagt tattaagtta tggagaccgg
     3421 tgtggcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3481 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccgg cctcagtgag
     3541 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     3601 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3841 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3901 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3961 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4021 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4081 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4141 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4201 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4261 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4321 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4381 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4441 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4501 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4561 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4621 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4681 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4741 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4801 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4861 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4921 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4981 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5041 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5101 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5161 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5221 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5281 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5341 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5401 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5461 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5521 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5581 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5641 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5701 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5761 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5821 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5881 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5941 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6001 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6061 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
