LOCUS       sequence-491776-        6611 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  3' AAVLINK plasmid for ZMYM2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  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  JOURNAL   Unpublished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 6611)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3'-ZMYM2-FLAG
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3319..3384
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3461..3481)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3874..4014
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3874..4003
                     /label=AAV2 ITR
     rep_origin      4089..4544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4176..4195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4386..4407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4561..4580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4680..4702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4740..4758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4826..4930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4931..4999,5000..5791)
                     /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(5149..5168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5962..6550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6451..6470
                     /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 gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtgactg ttgtaaatct
     1501 caaggaactc ttaaagagcg agttcagtgg cgtggggaaa tgaaacattt ctgtgatcaa
     1561 cattgcttac tgcgtttcta ctgtcaacaa aatgagccca acatgacaac tcagaaagga
     1621 cctgaaaact tacattatga tcagggttgt cagacatctc gaaccaaaat gacaggttca
     1681 gcaccacccc cttctccaac acctaacaaa gagatgaaga acaaagcagt tctttgcaaa
     1741 cctttaacaa tgacaaaagc tacttactgt aaacctcaca tgcagaccaa atcttgtcag
     1801 acagatgata cttggaggac agaatatgtt ccagtgccta tccctgtgcc tgtgtatatc
     1861 ccagttccta tgcacatgta cagtcagaat attcctgttc ctactacagt tcctgttcct
     1921 gtgccagttc ctgtttttct gcctgctcca ttggacagca gtgagaagat tcatgcagca
     1981 attgaggagc taaaaagcaa ggtttcttca gatgctcttg atacagagtt gcttacaatg
     2041 acggatatga tgagtgaaga cgaggggaaa acagagacaa ccaacatcaa cagtgtaatt
     2101 attgaaacag atataattgg ttcagacctt ttgaagaact ctgacccaga gacacagtcc
     2161 agcatgcctg atgtaccata tgaaccagat ttggatatcg aaatagattt tcccagagct
     2221 gctgaggagc ttgatatgga aaatgaattt ttattaccac ctgtttttgg cgaagaatat
     2281 gaggaacagc ccagacctcg atctaaaaaa aagggagcca agagaaaggc tgtatcagga
     2341 taccagtctc atgatgatag ttctgacaat tcagaatgca gctttccttt caaatatacg
     2401 tatggcgtaa atgcatggaa acactgggtc aaaactaggc aacttgatga agatcttctg
     2461 gtattagatg agttaaaatc ttctaaatca gtaaagttaa aagaggatct actctctcac
     2521 accacagctg agcttaacta tgggttagct cattttgtca atgagatccg acggccaaat
     2581 ggagagaatt atgcacctga cagcatctat tacctttgcc ttggaataca ggagtatttg
     2641 tgtggaagta atcgaaaaga caacatattt attgatcctg gataccaaac atttgagcaa
     2701 gaattgaata aaatactgcg aagctggcaa ccaagcatac ttccagatgg gtcaatattc
     2761 tctcgagttg aagaagacta tctctggagg ataaaacaac taggatcaca ctctccagta
     2821 gctcttctga atacactgtt ctactttaac actaagtatt ttggcctgaa aacagtggaa
     2881 caacacttaa gactttcctt tggcactgtg tttaggcatt ggaaaaaaaa tcctttaacg
     2941 atggaaaaca aagcgtgtct tcgataccaa gtgtcttcct tgtgtggaac agataatgaa
     3001 gataaaatta ctactggaaa aagaaaacat gaagatgatg agccagtatt tgaacaaatt
     3061 gaaaacacag ccaatccttc cagatgtcct gtgaaaatgt ttgaatgcta cttgtctaaa
     3121 agtccacaga atcttaatca gaggatggat gttttttatt tgcaaccaga atgctctagt
     3181 tctacagata gccctgtctg gtatacgtct acttcactgg accgaaacac cttggaaaat
     3241 atgcttgtac gggttcttct agtaaaagat atttatgata aagacaatta tgaactggat
     3301 gaagacacag actgtacaga ctacaaagac catgacggtg attataaaga tcatgacatc
     3361 gactacaagg atgacgatga caagtaagaa ttctgatgac tcgagataat caacctctgg
     3421 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3481 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3541 tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc gccgcctgcc
     3601 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgtttattt
     3661 gtgaaatttg tgatgctatt gctttatttg taaccatcta gctttatttg tgaaatttgt
     3721 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3781 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagatctgga
     3841 gaccggtgtg gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     3901 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3961 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4021 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4081 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4441 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4561 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4621 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4681 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4741 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5461 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5641 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6601 tgctcacatg t
//
