pAAVLINKv1-pCALM1-Cre-3'-MINK1-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-MINK1-FLAG                6606 bp DNA     circular SYN 25-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6606)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 12, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6606
                     /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..3319
                     /codon_start=1
                     /gene="MINK1"
                     /gene_synonym="B55; MAP4K6; MEKKK 6; MINK; YSK2; ZC3"
                     /product="misshapen-like kinase 1 isoform 3"
                     /EC_number="2.7.11.1"
                     /label=MINK1
                     /note="isoform 3 is encoded by transcript variant 3; 
                     misshapen/NIK-related kinase; GCK family kinase MINK; MEK 
                     kinase kinase 6; MAPK/ERK kinase kinase kinase 6; 
                     mitogen-activated protein kinase kinase kinase kinase 6"
                     /db_xref="CCDS:CCDS45588.1"
                     /db_xref="GeneID:50488"
                     /db_xref="HGNC:HGNC:17565"
                     /db_xref="MIM:609426"
                     /protein_id="NP_722549.2"
                     /translation="PPAQPPGPPNASSNPDLRRSDPGWERSDSVLPASHGHLPQAGSLE
                     RNRVGASSKLDSSPVLSPGNKAKPDDHRSRPGRPADFVLLKERTLDEAPRPPKKAMDYS
                     SSSEEVESSEDDEEEGEGGPAEGSRDTPGGRSDGDTDSVSTMVVHDVEEITGTQPPYGG
                     GTMVVQRTPEEERNLLHADSNGYTNLPDVVQPSHSPTENSKGQSPPSKDGSGDYQSRGL
                     VKAPGKSSFTMFVDLGIYQPGGSGDSIPITALVGGEGTRLDQLQYDVRKGSVVNVNPTN
                     TRAHSETPEIRKYKKRFNSEILCAALWGVNLLVGTENGLMLLDRSGQGKVYGLIGRRRF
                     QQMDVLEGLNLLITISGKRNKLRVYYLSWLRNKILHNDPEVEKKQGWTTVGDMEGCGHY
                     RVVKYERIKFLVIALKSSVEVYAWAPKPYHKFMAFKSFADLPHRPLLVDLTVEEGQRLK
                     VIYGSSAGFHAVDVDSGNSYDIYIPVHIQSQITPHAIIFLPNTDGMEMLLCYEDEGVYV
                     NTYGRIIKDVVLQWGEMPTSVAYICSNQIMGWGEKAIEIRSVETGHLDGVFMHKRAQRL
                     KFLCERNDKVFFASVRSGGSSQVYFMTLNRNCIMNW"
     misc_feature    1535..1540
                     /label=SDS
     misc_feature    1806..1811
                     /label=SDS
     misc_feature    1833..1838
                     /label=SDS
     misc_feature    2838..2843
                     /label=SDS
     CDS             3326..3391
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3395..3826
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3539..3544
                     /label=SDS
     repeat_region   3869..4009
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    3869..3998
                     /label=AAV2 ITR
     repeat_region   3870..4010
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4084..4539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4821..4925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4926..5786
                     /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      5957..6545
                     /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 gggccccctg ctcagccccc
     1501 tggcccgccc aacgcctcta gtaaccccga cctcaggagg agcgaccctg gctgggaacg
     1561 ctcggacagc gtccttccag cctctcacgg gcacctcccc caggctggct cactggagcg
     1621 gaaccgcgtg ggagcctcct ccaaactgga cagctcccct gtgctctccc ctgggaataa
     1681 agccaagccc gacgaccacc gctcacggcc aggccggccc gcagactttg tgttgctgaa
     1741 agagcggact ctggacgagg cccctcggcc tcccaagaag gccatggact actcgtcgtc
     1801 cagcgaggag gtggaaagca gtgaggacga cgaggaggaa ggcgaaggcg ggccagcaga
     1861 ggggagcaga gatacccctg ggggccgcag cgatggggat acagacagcg tcagcaccat
     1921 ggtggtccac gacgtcgagg agatcaccgg gacccagccc ccatacgggg gcggcaccat
     1981 ggtggtccag cgcacccctg aagaggagcg gaacctgctg catgctgaca gcaatgggta
     2041 cacaaacctg cctgacgtgg tccagcccag ccactcaccc accgagaaca gcaaaggcca
     2101 aagcccaccc tcgaaggatg ggagtggtga ctaccagtct cgtgggctgg taaaggcccc
     2161 tggcaagagc tcgttcacga tgtttgtgga tctagggatc taccagcctg gaggcagtgg
     2221 ggacagcatc cccatcacag ccctagtggg tggagagggc actcggctcg accagctgca
     2281 gtacgacgtg aggaagggtt ctgtggtcaa cgtgaatccc accaacaccc gggcccacag
     2341 tgagacccct gagatccgga agtacaagaa gcgattcaac tccgagatcc tctgtgcagc
     2401 cctttggggg gtcaacctgc tggtgggcac ggagaacggg ctgatgttgc tggaccgaag
     2461 tgggcagggc aaggtgtatg gactcattgg gcggcgacgc ttccagcaga tggatgtgct
     2521 ggaggggctc aacctgctca tcaccatctc agggaaaagg aacaaactgc gggtgtatta
     2581 cctgtcctgg ctccggaaca agattctgca caatgaccca gaagtggaga agaagcaggg
     2641 ctggaccacc gtgggggaca tggagggctg cgggcactac cgtgttgtga aatacgagcg
     2701 gattaagttc ctggtcatcg ccctcaagag ctccgtggag gtgtatgcct gggcccccaa
     2761 accctaccac aaattcatgg ccttcaagtc ctttgccgac ctcccccacc gccctctgct
     2821 ggtcgacctg acagtagagg aggggcagcg gctcaaggtc atctatggct ccagtgctgg
     2881 cttccatgct gtggatgtcg actcggggaa cagctatgac atctacatcc ctgtgcacat
     2941 ccagagccag atcacgcccc atgccatcat cttcctcccc aacaccgacg gcatggagat
     3001 gctgctgtgc tacgaggacg agggtgtcta cgtcaacacg tacgggcgca tcattaagga
     3061 tgtggtgctg cagtgggggg agatgcctac ttctgtggcc tacatctgct ccaaccagat
     3121 aatgggctgg ggtgagaaag ccattgagat ccgctctgtg gagacgggcc acctcgacgg
     3181 ggtcttcatg cacaaacgag ctcagaggct caagttcctg tgtgagcgga atgacaaggt
     3241 gttttttgcc tcagtccgct ctgggggcag cagccaagtt tacttcatga ctctgaaccg
     3301 taactgcatc atgaactggt gtacagacta caaagaccat gacggtgatt ataaagatca
     3361 tgacatcgac tacaaggatg acgatgacaa gtaactcgag ataatcaacc tctggattac
     3421 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3481 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3541 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     3601 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa
     3661 atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa tttgtgatgc
     3721 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3781 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagat ctggagaccg
     3841 gtgtggcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     3901 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3961 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4021 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4081 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4141 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4261 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4321 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4381 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4441 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4501 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4561 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4621 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4681 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4741 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4801 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4861 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4921 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4981 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5041 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5101 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5161 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5221 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5281 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5341 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5401 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5461 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5521 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5581 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5641 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5701 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5761 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5821 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6601 acatgt
//