pAAVLINKv1-pCALM1-Cre-ASH1L-C-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-ASH1L-C-FLAG               6710 bp DNA     circular SYN 10-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     promoter        145..265
                     /label=pCALM1
     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"
     intron          1309..1349
                     /label=Splicing Donor
     protein_bind    1356..1389
                     /label=lox2272-JTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          1390..1481
                     /label=Splicing Acceptor
                     /note="Splicing acceptor"
     CDS             1482..3414
                     /codon_start=1
                     /gene="ASH1L"
                     /gene_synonym="ASH1; ASH1L1; KMT2H; MRD52"
                     /product="histone-lysine N-methyltransferase ASH1L 
                     isoform2"
                     /EC_number="2.1.1.359; 2.1.1.367"
                     /label=ASH1L
                     /note="isoform 2 is encoded by transcript variant 
                     2;probable histone-lysine N-methyltransferase ASH1L; 
                     ASH1-like protein; lysine N-methyltransferase 2H; absent 
                     small and homeotic disks protein 1 homolog; ash1 (absent, 
                     small, or homeotic)-like"
                     /db_xref="GI:110349788"
                     /db_xref="CCDS:CCDS1113.2"
                     /db_xref="GeneID:55870"
                     /db_xref="HGNC:HGNC:19088"
                     /db_xref="MIM:607999"
                     /protein_id="NP_060959.2"
                     /translation="RGHLSEEPSENINTPTRLTPQLQMKPMSNRERNFVLKHHVFLVRN
                     WEKIRQKQEEVKHTSDNIHSASLYTRWNGICRDDGNIKSDVFMTQFSALQTARSVRTRR
                     LAAAEENIEVARAARLAQIFKEICDGIISYKDSSRQALAAPLLNLPPKKKNADYYEKIS
                     DPLDLITIEKQILTGYYKTVEAFDADMLKVFRNAEKYYGRKSPVGRDVCRLRKAYYNAR
                     HEASAQIDEIVGETASEADSSETSVSEKENGHEKDDDVIRCICGLYKDEGLMIQCDKCM
                     VWQHCDCMGVNSDVEHYLCEQCDPRPVDREVPMIPRPHYAQPGCVYFICLLRDDLLLRQ
                     GDCVYLMRDSRRTPDGHPVRQSYRLLSHINRDKLDIFRIEKLWKNEKEERFAFGHHYFR
                     PHETHHSPSRRFYHNELFRVPLYEIIPLEAVVGTCCVLDLYTYCKGRPKGVKEQDVYIC
                     DYRLDKSAHLFYKIHRNRYPVCTKPYAFDHFPKKLTPKKDFSPHYVPDNYKRNGGRSSW
                     KSERSKPPLKDLGQEDDALPLIEEVLASQEQAANEIPSLEEPEREGATANVSEGEKKTE
                     ESSQEPQSTCTPEERRHNQRERLNQILLNLLEKIPGKNAIDVTYLLEEGSGRKLRRRTL
                     FIPENSFRK"
     misc_feature    1640..1645
                     /label=SDS
     misc_feature    3074..3079
                     /label=SDS
     CDS             3418..3483
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3499..3930
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3643..3648
                     /label=SDS
     repeat_region   3973..4113
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    3973..4102
                     /label=AAV2 ITR
     repeat_region   3974..4114
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4188..4643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4925..5029
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5030..5890
                     /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"
     rep_origin      6061..6649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ggagaggcca tctctctgag
     1501 gaacccagtg aaaatatcaa caccccaact agattgaccc cccaattaca gatgaagcca
     1561 atgtccaatc gtgaaaggaa ctttgtgtta aagcatcatg tattcttggt ccgaaactgg
     1621 gagaagattc gtcaaaaaca ggaggaagta aagcacacca gtgataatat tcactcagca
     1681 tcattatata cccgttggaa tgggatctgc cgagatgatg ggaatatcaa gtctgatgtc
     1741 ttcatgaccc agttctctgc cctgcagaca gctcgatctg ttcgaacaag acggttggca
     1801 gctgcagagg aaaatattga agtggctcgg gcagcccgcc tagcccagat cttcaaagaa
     1861 atttgtgatg gtatcatctc ttataaagat tcttcccggc aagcactggc agctccactt
     1921 ttgaaccttc ccccaaagaa aaagaatgct gattattatg agaagatctc tgatccccta
     1981 gatcttatca ccatagagaa gcagatcctc actggttact ataagacagt ggaagctttt
     2041 gatgctgaca tgctcaaagt ctttcggaat gctgagaagt actatgggcg taaatcccca
     2101 gttgggagag atgtttgtcg tctacgaaag gcctattaca atgcccggca tgaggcatca
     2161 gcccagattg atgagattgt gggagagaca gcaagtgagg cagacagcag tgagacctca
     2221 gtctctgaaa aggagaatgg gcatgagaag gacgacgatg ttattcgctg tatctgtggc
     2281 ctctacaagg atgaaggtct catgatccag tgtgacaagt gcatggtatg gcagcactgt
     2341 gattgtatgg gagtgaactc agatgtggag cactaccttt gtgagcagtg tgacccaagg
     2401 cctgtggaca gggaggttcc catgatccct cggccccact atgcccaacc tggctgtgtc
     2461 tacttcatct gtttgctccg agatgacttg ctgcttcgtc agggtgactg tgtgtatctg
     2521 atgagggata gtcggcgcac ccctgatggc cacccggtcc gtcagtccta tcgactgtta
     2581 tctcacatta accgagataa acttgacatc tttcgcattg agaagctttg gaagaatgaa
     2641 aaagaggaac ggtttgcctt tggtcaccat tatttccgtc cccacgaaac acaccactct
     2701 ccatcccgtc ggttctatca taatgaacta tttcgggtgc cactctatga gatcattccc
     2761 ttggaggctg tagtggggac ctgctgtgtg ttggaccttt atacgtattg taaagggaga
     2821 cccaaaggag taaaggagca agatgtgtac atctgtgatt atcggcttga caagtcagca
     2881 cacctgtttt acaagatcca ccggaaccgc tatcctgtct gcaccaaacc ctatgctttt
     2941 gatcacttcc ccaagaagct cactcccaaa aaagatttct cgcctcatta cgtcccagac
     3001 aactacaaga ggaatggagg acgatcatcc tggaagtctg agcgctcaaa gccaccccta
     3061 aaagacttgg gccaggagga tgatgctcta cccttgattg aagaggttct agccagtcaa
     3121 gagcaagcag ccaatgagat acccagcctg gaggagccag aacgggaagg ggccactgct
     3181 aacgtcagtg agggtgaaaa aaaaacagag gaaagtagtc aagaacccca gtcaacctgt
     3241 acccctgagg aacgacggca taaccaacgg gaacgactca accagatctt gctcaatctc
     3301 cttgaaaaaa tccctggaaa aaatgccatt gatgtgacct acttgctgga ggaaggatca
     3361 ggcaggaaac tgcgaaggcg tactttgttt atcccagaaa acagctttcg aaagaaggac
     3421 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3481 aagtaagaat tctgatgact cgagataatc aacctctgga ttacaaaatt tgtgaaagat
     3541 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3601 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3661 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3721 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     3781 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3841 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3901 gttcaggggg agatgtggga ggttttttaa agatctggag accggtgtgg cacgtgcgga
     3961 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4021 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4081 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     4141 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     4201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4261 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4381 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4501 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4561 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4621 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4681 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4741 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4801 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4861 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4921 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4981 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5041 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5101 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5161 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5221 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5281 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5341 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5401 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5461 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5521 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5581 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5641 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5701 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5761 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5821 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5881 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5941 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6001 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6061 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6121 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6181 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6241 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6301 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6361 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6421 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6481 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6541 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6601 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6661 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//