LOCUS       sequence-476876-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for POGZ expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  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  JOURNAL   Unpublished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 6168)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3' POGZ
FEATURES             Location/Qualifiers
     source          1..6168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1674..1739
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1816..1836)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2229..2369
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2229..2358
                     /label=AAV2 ITR
     rep_origin      2444..2899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2741..2762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2916..2935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3035..3057
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3095..3113)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3181..3285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3286..3354,3355..4146)
                     /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(3504..3523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4317..4905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4806..4825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4967..5096
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(5243..6168,1..106)
                     /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"
ORIGIN
        1 catgcaagct ggtggctgga ccaatgtaaa tattgtaatg aactatatcc gtaacctgga
       61 tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gattaggtaa gtatcaaggt
      121 tacaagacag gtttaaggag accaataact agtataactt cgtataggat actttatagc
      181 aatttatgaa actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta
      241 ttggtcttac tgacatccac tttgcctttc tctccacagg gagcccagtc acccaagaac
      301 ctgagctagc atcaggtggt ggtggtagtg gtggagttgg caaaaaggag cagctgtctg
      361 tgaagaagct tcgagtagta ctgtttgctc tatgctgcaa tacagaacag gcagctgaac
      421 acttccgaaa tccccagcga cgtattcgcc gttggcttcg acgtttccag gcctcccagg
      481 gggagaatct agagggcaaa tatctgagct ttgaggcaga agagaaactg gctgagtggg
      541 tgctaaccca gcgcgaacaa cagctacctg taaatgagga gaccttgttc cagaaggcca
      601 ccaaaatagg acgttctttg gaaggggggt ttaagatctc ctatgagtgg gctgtgcgtt
      661 tcatgctgcg gcaccacctg actccccatg cccggcgagc tgtggcccac accctaccta
      721 aggatgtagc agagaatgca ggactcttca ttgattttgt acaacggcag attcacaacc
      781 aggacttacc cttgtctatg attgtggcta ttgatgagat ctctttgttc ctggatacag
      841 aggtgctgag cagtgatgat cgaaaggaga atgccctgca gacagtgggc acaggggaac
      901 cttggtgtga tgtagtccta gccattctgg cagatggcac tgtccttccc accctggttt
      961 tctacagagg gcagatggat cagcctgcta acatgccaga ctccatattg ctagaggcaa
     1021 aggagagtgg ctacagtgat gacgagatca tggagctgtg gtcaactcga gtgtggcaga
     1081 agcacacagc ttgccagcgc agcaaaggca tgcttgtgat ggactgtcat cgcactcact
     1141 tgtcagaaga ggtactggct atgcttagtg cctctagcac tttgcctgca gtggtcccag
     1201 caggctgtag ctccaaaatt cagccattag atgtatgcat caaaagaact gtcaagaact
     1261 tcctgcataa aaaatggaag gaacaggctc gggaaatggc agatactgca tgtgattctg
     1321 atgtcctgct tcagctggtg cttgtctggc tgggtgaagt gctaggtgtc attggggact
     1381 gtccagagct agttcagcgc tccttcctgg tggctagtgt tctgcctggc cccgatggca
     1441 acattaactc acctacaaga aatgctgaca tgcaggagga gctaattgcc tccctagagg
     1501 agcaactgaa gctgagtggg gaacattctg agtcttccac tccacgaccc agatcatctc
     1561 ctgaagagac aattgagcct gaaagtcttc accagctctt tgagggtgaa agtgagaccg
     1621 agtctttcta tggctttgaa gaagctgacc tagatctgat ggagatttgt acagactaca
     1681 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     1741 aagaattctg atgactcgag ataatcaacc tctggattac aaaatttgtg aaagattgac
     1801 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1861 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1921 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1981 gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt
     2041 atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2101 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2161 agggggagat gtgggaggtt ttttaaagat ctggagaccg gtgtggcacg tgcggaccga
     2221 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     2281 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     2341 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     2401 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     2461 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2521 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2581 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2641 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2701 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2761 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2821 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2881 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     2941 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3001 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3061 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3121 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3241 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3301 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3361 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3421 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3481 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3541 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3601 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3661 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3721 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3781 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3841 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3901 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3961 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4021 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4081 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4141 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4201 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4261 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4321 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4381 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4441 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4501 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4561 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4621 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4681 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4741 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4801 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4861 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4921 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
     4981 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
     5041 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     5101 ccgcacgcgt cgctctgcgc gaggcggctc cgccgcggcg gagggatacg gcgcaccata
     5161 tatatatcgc ggggcgcaga ctcgcgctcc ggcagtggtg ctgggagtgt cgtggacgcc
     5221 gtgccgttac tcgtacgcca ccatgtccaa tttactgacc gtacaccaaa atttgcctgc
     5281 attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga
     5341 tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc
     5401 ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga
     5461 ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa accatccagc aacatttggg
     5521 ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt
     5581 ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca
     5641 ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga
     5701 tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg
     5761 tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat
     5821 gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact
     5881 tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga
     5941 tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac
     6001 cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta
     6061 cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt
     6121 cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagat
//
