LOCUS       sequence-491422-        6593 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for SETD1A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6593)
  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  JOURNAL   Unpublished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 6593)
  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            "Unpublished"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'-SETD1A-FLAG
FEATURES             Location/Qualifiers
     source          1..6593
                     /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"
     CDS             3313..3378
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3443..3463)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3856..3996
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3856..3985
                     /label=AAV2 ITR
     rep_origin      4071..4526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4158..4177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4368..4389
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4543..4562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4662..4684
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4722..4740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4808..4912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4913..4981,4982..5773)
                     /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(5131..5150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5944..6532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6433..6452
                     /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 gggttgcagg ctccccagtc
     1501 acacccctgc ccgaacagga ggcgtctcca gcaaggcctg caggccccac ggaggagtca
     1561 ccccccagtg cgcctctgcg tcccccagaa ccacctgctg ggcccccggc ccctgcccca
     1621 cgccccgatg agcgtccctc ttctcccatc cccctcctgc ccccacccaa gaaacgccgg
     1681 aaaactgtct ccttctctgc catcgaggtg gtgccagccc cggagccccc tccagccaca
     1741 ccgccgcagg ccaagtttcc cggcccagcc tcccgcaagg ctccccgggg cgtggagcgg
     1801 accatccgca acctgcccct ggaccacgca tctctggtca agagttggcc cgaggaggtg
     1861 tcccgaggag gccggagccg ggctggaggc cgaggccgcc tcaccgagga agaggaggct
     1921 gagccaggga cagaggtgga cctggcggtc ctggccgacc tggccctgac ccctgcccgg
     1981 cgcgggctgc ctgccctgcc tgctgttgaa gactcagagg ccacagagac atcggacgag
     2041 gccgagcgcc ctaggcccct gctcagccac atcctcctgg agcacaacta tgccctggcc
     2101 gtcaagccca cgccccctgc gccagccctg cggcccccgg agccagtgcc cgcacccgcc
     2161 gccctcttca gttccccagc tgatgaggtc ctggaggccc ccgaggtggt ggtggctgag
     2221 gcggaggagc ccaagccgca gcaactgcag cagcagcggg aggagggcga agaggagggg
     2281 gaggaagagg gggaggaaga ggaggaggag tcctctgaca gcagcagcag cagcgatggg
     2341 gagggcgccc tccggaggcg cagcctccgc tcccacgccc ggcgccgccg ccctccgccc
     2401 ccacccccgc cgccaccgcc ccgcgcctac gagccacgca gtgagtttga acagatgacc
     2461 atcctgtatg acatttggaa ctcgggcctg gactcagagg acatgagtta cctgcggctt
     2521 acgtacgagc ggctgctgca gcagacaagc ggggctgact ggctcaacga cactcactgg
     2581 gtccatcaca caatcaccaa cctgaccacc ccaaaacgca agcggcggcc ccaggatggg
     2641 ccccgggagc accagacagg ctcagcccgc agcgaaggct actaccccat cagcaagaag
     2701 gagaaggaca agtacctgga cgtgtgccca gtctcggccc ggcagctgga gggcgtggac
     2761 actcagggga cgaaccgcgt gctgtccgag cgccggtccg agcagcggcg gctgctgagc
     2821 gccatcggta cctccgccat catggacagt gacctgctga aactcaacca gctcaagttc
     2881 cggaagaaga agctccgatt tggccggagc cggatccacg agtggggtct gtttgccatg
     2941 gaacccattg ctgctgacga gatggtcatc gaatacgtgg gtcagaacat ccgtcagatg
     3001 gtggccgaca tgcgggagaa gcgctacgtg caggagggca ttggcagcag ctacctgttc
     3061 cgggtggacc acgacaccat catcgatgcc accaagtgtg gcaacctggc cagattcatc
     3121 aaccactgct gcacgcctaa ctgctacgcc aaggtcatca ccatcgagtc ccagaagaag
     3181 atcgtgatct actccaagca gcccattggc gtggacgagg agatcaccta cgactacaag
     3241 ttcccactgg aagacaacaa gatcccgtgt ctgtgtggca cagagagctg ccggggctcc
     3301 ctaaactgta cagactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3361 aaggatgacg atgacaagta actcgagata atcaacctct ggattacaaa atttgtgaaa
     3421 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3481 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3541 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3601 gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta
     3661 ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt
     3721 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3781 caggttcagg gggagatgtg ggaggttttt taaagatctg gagaccggtg tggcacgtgc
     3841 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3901 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3961 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4021 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4081 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4141 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4201 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4261 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4321 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4381 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4441 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4501 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4561 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4621 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4681 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4741 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4801 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4861 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4921 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4981 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5041 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5101 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5161 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5221 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5281 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5341 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5401 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5461 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5521 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5581 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5641 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5701 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5761 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
