LOCUS       sequence-479590-        5697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for NEO1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5697)
  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  JOURNAL   Unpublished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 5697)
  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            "Unpublished"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-5' NEO1
FEATURES             Location/Qualifiers
     source          1..5697
                     /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"
     repeat_region   2960..3100
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2960..3089
                     /label=AAV2 ITR
     rep_origin      3175..3630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3262..3281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3472..3493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3647..3666)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3766..3788
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3826..3844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4017..4085,4086..4877)
                     /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(4235..4254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5537..5556
                     /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 ctggatctgc ggcggagcgg ggagcccggc gactcctcag
      781 caccccctcc ttctggctct actgcctgct gctgctcggg cgccgggcgc cgggcgccgc
      841 ggccgccagg agcggctccg cgccgcagtc cccaggagcc agcattcgaa cgttcactcc
      901 attttatttt ctggtggagc cggtggatac actctcagtt agaggctctt ctgttatatt
      961 aaactgttca gcatattctg agccttctcc aaaaattgaa tggaaaaaag atggaacttt
     1021 tttaaactta gtatcagatg atcgacgcca gcttctcccg gatggatctt tatttatcag
     1081 caatgtggtg cattccaaac acaataaacc tgatgaaggt tattatcagt gtgtggccac
     1141 tgttgagagt cttggaacta ttatcagtag aacagcgaag ctcatagtag caggtcttcc
     1201 aagatttacc agccaaccag aaccttcctc agtttatgct gggaacaatg caattctgaa
     1261 ttgtgaagtt aatgcagatt tggtcccatt tgtgaggtgg gaacagaaca gacaacccct
     1321 tcttctggat gatagagtta tcaaacttcc aagtggaatg ctggttatca gcaatgcaac
     1381 tgaaggagat ggcgggcttt atcgctgcgt agtggaaagt ggtgggccac caaagtatag
     1441 tgatgaagtt gaattgaagg ttcttccaga tcctgaggtg atatcagact tggtattttt
     1501 gaaacagcct tctcccttag tcagagtcat tggtcaggat gtagtgttgc catgtgttgc
     1561 ttcaggactt cctactccaa ccattaaatg gatgaaaaat gaggaggcac ttgacacaga
     1621 aagctctgaa agattggtat tgctggcagg tggtagcctg gagatcagtg atgttactga
     1681 ggatgatgct gggacttatt tttgtatagc tgataatgga aatgagacaa ttgaagctca
     1741 agcagagctt acagtgcaag ctcaacctga attcctgaag cagcctacta atatatatgc
     1801 tcacgaatct atggatattg tatttgaatg tgaagtgact ggaaaaccaa ctccaactgt
     1861 gaagtgggtc aaaaatgggg atatggttat cccaagtgat tattttaaga ttgtaaagga
     1921 acataatctt caagttttgg gtctggtgaa atcagatgaa gggttctatc agtgcattgc
     1981 tgaaaatgat gttggaaatg cacaagctgg agcccaactg ataatccttg aacatgcacc
     2041 agccacaacg ggaccactgc cttcagctcc tcgggatgtc gtggcctccc tggtctctac
     2101 ccgcttcatc aaattgacgt ggcggacacc tgcatcagat cctcacggag acaaccttac
     2161 ctactctgtg ttctacacca aggaagggat tgctagggaa cgtgttgaga ataccagtca
     2221 cccaggagag atgcaagtaa ccattcaaaa cctaatgcca gcgaccgtgt acatctttag
     2281 agttatggct caaaataagc atggctcagg agagagttca gctccactgc gagtagaaac
     2341 acaacctgag gttcagctcc ctggcccagc acctaacctt cgtgcatatg cagcttcgcc
     2401 tacctccatc actgttacgt gggaaacacc agtgtctggc aatggggaaa ttcagaatta
     2461 taagttgtac tacatggaaa aggggactga taaagaacag gatgttgatg tttcaagtca
     2521 ctcttacacc attaatgggt tgaaaaaata tacagagtat agtttccgag tggtggccta
     2581 caataaacat ggtcctggag tttccacacc agatgttgct gttcgaacat tgtcagatgt
     2641 tcccagtgct gctcctcaga atctgtcctt ggaagtgaga aattcaaaga gtattatgat
     2701 tcactggcag ccacctgctc cagccacaca aaatgggcag attactggct acaagattcg
     2761 ctaccgaaag gcctcccgaa agagtgatgt cactgagacc ttggtaagcg ggacacagct
     2821 gtctcagctg attgaaggta agtatcaagg ttacaagaca ggtttaagga gaccaataag
     2881 ctattattcg tataggatac tttatacgaa gttattaagt tatggagacc ggtgtggcac
     2941 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     3001 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     3061 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     3121 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     3181 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3241 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3301 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3361 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3421 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3481 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     3541 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3601 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3661 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3721 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3781 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
