LOCUS       sequence-491404-        6148 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for SETBP1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  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  JOURNAL   Unpublished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 6148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 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            "Unpublished"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'-SETBP1-FLAG
FEATURES             Location/Qualifiers
     source          1..6148
                     /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             2868..2933
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2998..3018)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3411..3551
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3411..3540
                     /label=AAV2 ITR
     rep_origin      3626..4081
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3713..3732)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3923..3944
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4098..4117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4217..4239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4277..4295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4363..4467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4468..4536,4537..5328)
                     /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(4686..4705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5499..6087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5988..6007
                     /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 ggtaggcagt gccagtctgt
     1501 ccagtggtcg gctccataag aggaaacaca aacacaagca taagcacaag gaagaccgga
     1561 tcctagggac ccatgacaac ctgagtggtc tttttgcagg caaagccaca ggcttctcca
     1621 gccacatcct gagcgagcgg ctgagtagcg cagacaaaga gctcccgctg gtgagtgaga
     1681 agaacaagca taaggagaaa cagaagcacc agcacagcga agccggccac aaagcttcta
     1741 agaacaactt tgaggtggac accctgtcta cactgtcact ttccgacgcc cagcattgga
     1801 cacaggccaa ggaaaaagga gacttgagca gtgagcctgt ggactcatgc acgaaaagat
     1861 actctggcag tggcggggat ggtggcagca cgagatcgga gaacctggac gtgttcagtg
     1921 aaatgaaccc ttcgaatgac aagtgggaca gtgacgtgag tgggagtaaa aggaggagct
     1981 atgaaggctt tggaacgtac agggaaaagg acatccaagc cttcaagatg aaccgcaagg
     2041 agagaagttc ttatgactcc tccatgtctc cagggatgcc aagtccccac ttaaaagtgg
     2101 accagacagc agtgcatagt aagaacgaag gctcagtgcc caccatgatg accaggaaga
     2161 agccagccgc agttgacagt gttacaattc caccagcccc agtgttatct ctccttgctg
     2221 catctgcagc aacgtcggat gcagtcggct cctccctgaa gaagaggttc aagcggcggg
     2281 agatcgaagc catccagtgc gaagtgcgga agatgtgcaa ctacaccaag atcctgtcca
     2341 ccaagaagaa cctggaccac gtgaacaaga tcctgaaggc caagcggctg cagagacaat
     2401 caaaaacagg caacaacttc gtgaagaaga ggcgcgggcg tcccaggaag cagcccaccc
     2461 agttcgatga ggactccaga gaccaaatgc cggtgctgga aaaatgcatc gacctgccca
     2521 gcaaaagagg ccagaagccc agcctgagcc cgctggtgct ggagcccgcc gccagccaag
     2581 acaccatcat ggccaccatc gaggcggtca tccacatggc ccgggaggcg ccgcccctgc
     2641 ccccgccacc gccgccgccc ctgccgccac cgccgccacc acccctgccc ccgccacccc
     2701 ctctacccaa gaccccccga ggcggaaaga ggaaacacaa accgcaggcc cccgctcagc
     2761 ccccacagca gtcgcccccg cagcagcccc ttccccagga agaggaggtg aaagccaaaa
     2821 ggcagaggaa gtcccgaggg agtgagagcg aggtccttcc ctgtacagac tacaaagacc
     2881 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtaactcg
     2941 agataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3001 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3061 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     3121 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3181 ttccgtggtg tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccatctagct
     3241 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3301 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3361 ttttttaaag atctggagac cggtgtggca cgtgcggacc gagcggccgc aggaacccct
     3421 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3481 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3541 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3601 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3661 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3721 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3781 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3841 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3901 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3961 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4021 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4081 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4141 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4201 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4261 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4321 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4381 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4441 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4501 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4561 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4621 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4681 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4741 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4801 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4861 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4921 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4981 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5041 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5101 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5161 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5221 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5281 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5341 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5401 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5641 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6121 gccttttgct ggccttttgc tcacatgt
//
