LOCUS       sequence-489086-        5893 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for ZMYM2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  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  JOURNAL   Unpublished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 5893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 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            "Unpublished"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'-ZMYM2
FEATURES             Location/Qualifiers
     source          1..5893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2235..2261)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2298..2566
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2579..2790)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   2835..2964
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3026..3614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3106..3125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3785..4576,4577..4645))
                     /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     4408..4427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4646..4750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4818..4836
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4874..4896)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4996..5015
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5032..5487)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5169..5190)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5381..5400
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5562..5702
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5573..5702
                     /label=AAV2 ITR
ORIGIN
        1 ccatcgagtt ctttagttgc tcccttctta catagctgag aacaatagtt gcaagtaaca
       61 catctcaatc ctaaacgtct ggcaaaatcc tgtttgtata ataatttgca gccttcacta
      121 cagaaaggtc tcttaacgcc agagaaattt actgtttcat gaagagtctt ctcctcttga
      181 cagtattcgc aatatgtaac tatgcaatgc aacttcttat agtcatctga acaagttttg
      241 ctgcagaact gatgcacttt gttctcccat tccaggattt ctggttttga acaaaaggaa
      301 tttttgcagt agttgcattt caactgaata tcacttggcg ctacaaactg gccatttgga
      361 gatgactgca tacttagagc ctgaaattta gccacacaac tggaattgca aaagttgtac
      421 agttttccat caggcattgt ggcttggtat tgaggtaaag agtttcgctt acaaaaatgg
      481 caaataattc ccaaactttg tgattttgca tattttgtct tgtgactgtt catacaagca
      541 gttgaacaat atggctccat atatccatta ggacctatac actgagtcat atcaaaaaac
      601 ctgcactgtg ttcggcaacc agtacaagtt gtcagttttc catatttctc aggctgcatt
      661 aagaaagagt cctgcatgtg atctcgaacc tccttcagga agcttggatg gctacctacc
      721 tgtttgtatt cactgacaca actttgacag caaaatcttt tctgttgacc atcaatcacc
      781 agaacattat ttccagcacc tttactgggc aagtactctc cacactgttc acagcaattc
      841 attattaaac cattggccat tctatatcta ttaaagcagt ggtcactgca cagcttatga
      901 gtcatatttt taaagctgac ttcatggcga atctcagtta gtttaccaca gattgtacat
      961 cttgatttat ttagagctcc tttagtagga ttctgtttgt cttcatagag agataaacaa
     1021 gatgtactac agaattcctg gaaggactca cttgaatcca cttgagcaac aatggttcct
     1081 ttcattgtag ttatatcttt tttacacata acacagagtt tctttggagc aggcttgtgg
     1141 gagaaggaag aaaggcaggt ggtagaacaa aagaggtgag ctgatccttt tcgttgataa
     1201 gctgtctggc ccttctgtaa aggttttttg cagtttgcac aagtgacttt aactggttta
     1261 gtaggctgct gttggacaga cggctggaaa atctgtttag gaagtgaggc cactggtgat
     1321 aaagagtcca cccctggttg tttctgatta cggggaaatg aagctgactg ggagatccaa
     1381 gaatcaggat tatgatgagt tgcagattct ccattctgaa aaacgtctcc tgccacattc
     1441 attctaccag gattaaaagg tcctactcca gtcttggtct gtgaagttaa agatggggtg
     1501 tatgtttgta tattaacacc aaaattactt gactgcagtc cgttagagac atctcccaag
     1561 ttggtattct gcagtgatgt tacatgtgta atcattaagt tcatatctcg cccgtcagta
     1621 ttttctaatt ggccatcatt cacagagctt acacctgttt ctaaagttgt aacattagca
     1681 atctgaattt cagagtctgg ttctgtggtg ataccactat tacccattcc tgcatttacc
     1741 ttacttcttg aaaaactgga agtggagaaa tccacatcat tggttctgtt tttagtctca
     1801 ggaggtcttc gttcaataaa attggaggaa tttttctctt gcccttgatt tgtttccatg
     1861 tcctcttcat catcaatgac aattgtctca cttacacttc ccttctgaga tgccaactct
     1921 tttgaggaag gagtaatttt ggaatcattt ccttgtagtt cttcattttt tgatgatgta
     1981 aatgttatgg ttctttgatc agctaccact ggtacagaag gtgggggagg ttgtacaggt
     2041 tcgataaaaa caacatcatc atcatcttcc actgacgagt tctgaaactt attagatcta
     2101 gacactaaag gattagctgg accactaaat gagtttccta cattcgtgag actagttgcc
     2161 atggccgtac tccctaataa aacaggagtc tgatcagtca attctaatcc tcccactgaa
     2221 cttgtgtcag atccagcata gtcaggtacg tcataagggt acatggtggc cgtacggtag
     2281 gcgccggtca cagcttggat ctgtaacggc gcagaacaga aaacgaaaca aagacgtaga
     2341 gttgagcaag cagggtcagg caaagcgtgg agagccggct gagtctaggt aggctccaag
     2401 ggagcgccgg acaaaggccc ggtctcgacc tgagctttaa acttacctag acggcggacg
     2461 cagttcagga ggcaccacag gcgggaggcg gcagaacgcg actcaaccgg cgtggatggc
     2521 ggcctcaggt agggcggcgg gcgcgtgaag gagagatgcg agcccctcga agcttcagct
     2581 gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat
     2641 acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt
     2701 ttaccccgcg ccaccttctc taggcacccg ttcaattgcc gacccctccc cccaacttct
     2761 cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcga tcgcagatcc
     2821 acgcgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2881 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg
     2941 agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3001 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3061 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3121 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3181 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3241 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3301 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3361 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3421 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3481 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3541 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3601 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3661 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3721 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3781 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3961 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4201 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4441 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4621 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4681 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4741 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4801 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4861 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4921 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4981 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa
     5041 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5101 aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga
     5161 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5221 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     5281 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5341 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5401 cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5461 cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat
     5521 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc
     5581 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     5641 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     5701 ctgcggccgc tcggtccgca cgtgccacac cggtctccat aacttaataa cttcgtataa
     5761 agtatcctat acgaataata gcttattggt ctccttaaac ctgtcttgta accttgatac
     5821 ttaccttgca gccttgtagt accaatcctg aaatttttta cagcaatctt cactgcagaa
     5881 atctctcaca aca
//
