LOCUS       sequence-476887-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for ROBO2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  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  JOURNAL   Unpublished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 6027)
  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            "Unpublished"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' ROBO2
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2287..2427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2287..2416
                     /label=AAV2 ITR
     rep_origin      2502..2957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2589..2608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2799..2820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2974..2993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3093..3115
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3153..3171)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3239..3343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3344..3412,3413..4204)
                     /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(3562..3581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4375..4963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4864..4883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5025..5154
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5199..5410
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5423..5691
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5728..5754
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 ttattcttct tgcgcatcgt gcacgggcgc aggagtaaac ctgatgaagg aagctacgtt
       61 tgtgttgcga ggaactatct tggtgaagca gtgagtcgaa atgcgtctct ggaagtggca
      121 ttgttacgag atgacttccg acaaaacccc acagatgttg tagtggcagc tggagagcct
      181 gcaatcctgg agtgccagcc tccccgggga cacccagaac ccaccatcta ctggaaaaaa
      241 gacaaagttc gaattgatga caaggaagaa agaataagta tccgtggtgg aaaactgatg
      301 atctccaata ccaggaaaag tgatgcaggg atgtatactt gtgttggtac caatatggtg
      361 ggagaaaggg acagtgaccc agcagagctg actgtctttg aacgacccac atttctcagg
      421 aggccaatta accaggtggt actggaggaa gaagctgtag aatttcgttg tcaagtccaa
      481 ggagatcctc aaccaactgt gaggtggaaa aaggatgatg cagacttgcc aagaggaagg
      541 tatgacatca aagacgatta cacactaaga attaaaaaga ccatgagtac agatgaaggc
      601 acctatatgt gtattgctga gaatcgggtt ggaaaaatgg aagcctctgc tacactcacc
      661 gtccgagctc gccctgttgc tcccccacag tttgtggttc ggccaagaga tcagattgtt
      721 gctcaaggtc gaacagtgac atttccctgt gaaactaaag gaaacccaca gccagctgtt
      781 ttttggcaga aagaaggcag ccagaaccta cttttcccaa accaacccca gcagcccaac
      841 agtagatgct cagtgtcacc aactggagac ctcacaatca ccaacattca acgttccgac
      901 gcgggttact acatctgcca ggctttaact gtggcaggaa gcattttagc aaaagctcaa
      961 ctggaggtta ctgatgtttt gacagataga cctccaccta taattctaca aggcccagcc
     1021 aaccaaacgc tggcagtgga tggtacagcg ttactgaaat gtaaagccac tggtgatcct
     1081 cttcctgtaa ttagctggtt aaaggaggga tttacttttc cgggtagaga tccaagagca
     1141 acaattcaag agcaaggcac actgcagatt aagaatttac ggatttctga tactggcact
     1201 tatacttgtg tggctacaag ttcaagtgga gagacttcct ggagtgcagt gctggatgtg
     1261 acagagtctg gagcaacaat cagtaaaaac tatgatttaa gtgacctgcc agggccacca
     1321 tccaaaccgc aggtcactga tgttactaag aacagtgtca ccttgtcctg gcagccaggt
     1381 acccctggaa cccttccagc aagtgcatat atcattgagg ctttcagcca atcagtgagc
     1441 aacagctggc agaccgtggc aaaccatgta aagaccaccc tctatactgt aagaggactg
     1501 cggcccaata caatctactt attcatggtc agagcgatca acccccaagg tctcagtgac
     1561 ccaagtccca tgtcagatcc tgtgcgcaca caagatatca gcccaccagc acaaggagtg
     1621 gaccacaggc aagtgcagaa agagctagga gatgtccttg tccgtcttca taatccagtt
     1681 gtgctgactc ccaccacggt tcaggtcaca tggacggttg atcgccaacc ccagtttatc
     1741 caaggctacc gagtgatgta tcgtcagact tcaggtctgc aggcgacatc ttcgtggcag
     1801 aatttagatg ccaaagtccc gactgaacga agtgctgtct tagtcaacct gaaaaagggg
     1861 gtgacttatg aaattaaagt acggccatat tttaatgagt tccaaggaat ggatagtgaa
     1921 tctaaaacgg ttcgtactac tgaagaagcc ccaagtgccc caccacagtc tgtcactgta
     1981 ctgacagttg gaagctacaa tagcacaagt attagtgttt cctgggatcc tcctcctcca
     2041 gatcaccaga atggaattat ccaagaatac aagatctggt gtctaggaaa tgaaacgcga
     2101 ttccatatca acaaaactgt ggatgcagcc attcggtccg taataattgg tggattattc
     2161 ccaggtaagt atcaaggtta caagacaggt ttaaggagac caataagcta ttattcgtat
     2221 aggatacttt atacgaagtt attaagttat ggagaccggt gtggcacgtg cggaccgagc
     2281 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2341 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2401 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2461 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2521 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2581 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2641 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2701 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2761 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2821 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2881 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2941 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3001 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3061 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3121 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3181 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3241 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3301 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3361 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3421 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3481 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3541 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3601 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3661 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3721 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3781 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3841 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3901 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3961 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4021 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4081 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4141 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4201 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4261 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4321 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4381 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4441 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4501 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4561 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4621 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4681 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4741 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4801 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4861 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4921 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4981 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     5041 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     5101 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     5161 gcacgcgtgg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     5221 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg
     5281 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     5341 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     5401 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac
     5461 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc
     5521 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc
     5581 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct
     5641 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg
     5701 accggcgcct accgtacggc caccatgtac ccttatgacg tacctgacta tgctggatct
     5761 atgagtctgc tgatgtttac acaactactg ctctgtggat ttttatatgt tcgggttgat
     5821 ggatcgcgtc ttcgccagga ggactttccc ccgcggattg tggagcatcc ttccgatgtc
     5881 atcgtctcta agggcgagcc cacgactctg aactgcaagg cggagggccg gccaacgccc
     5941 accattgagt ggtacaaaga tggggagcga gtggagactg acaaggacga tccccggtcc
     6001 cacaggatgc ttctgcccag cggatcc
//
