pAAVLINKv1-pCALM1-Cre-APC-C-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-APC-C-FLAG                6786 bp DNA     circular SYN 21-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6786
                     /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"
     misc_feature    12..130
                     /label=AAV2 ITR
     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..3499
                     /codon_start=1
                     /label=APC
                     /translation="GRKVHWKLKR*NLKVKESKEEKKFIKV*LLEKFDLIQKFQAK*NS
                     PFKQTCLQSLEAGQ*FIFQEFEIAPQVQVLFLKKAHPLRLQPPKALVKVKQPPLLLEEP
                     SHL*NQN*ALLPGRHPK*VGQVKHLLDQDLEIRPLQDLPSNH*VDLYSLLAETQFPLVE
                     ME*VLLTNYLNFQGHHPLVLLQLSPQVLEKCHIHLQVDR*ANRTLPNKQVYPRMPVVFQ
                     EVSLPPKD*IR*IMVMEPIKR*NFLECLQLNQVEVNLIDQKDLY*YASQLSSKKLQAQP
                     *EENWRNLLHLNLFLHHLDQLLPLGPRHKLQF*VLPFLICLYPHIRLFRLVDGENSHLI
                     SVPL*SIMMEDQQSAMILHGLILKVLLDFQSIGQEPGNVSTANIHHPFLE*ALGEELEV
                     HLQFFLLHQNPVKKQKVRMKNM*TLFQEPNKVKKTKYPQKEHGEK*KKMNFLPQIVLLR
                     PFPQVLQMVLNQRL*FIKWHLLFLKQRMFG*ELRTVPLTILDLEDLPQVILPR*LTVFQ
                     KRQIQTLKIQKIIRQNKMWVMAVFPCVPWVWKIA*TPLFRWMPLTKKELR*NQDKIILS
                     LYQRLMKVL*WNVPHSVLAAQANTVHLVGLLLPE*LLLITTQALGKAAQIALQLGHLRS
                     QLQ*ITTQRSEIPKLTAQNPVEPKVLSAILGLTL*HL"
     misc_feature    1537..1542
                     /label=SDS
     misc_feature    1982..1987
                     /label=SDS
     CDS             3506..3571
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3575..4006
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3719..3724
                     /label=SDS
     repeat_region   4049..4189
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4049..4178
                     /label=AAV2 ITR
     rep_origin      4264..4719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5001..5105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5106..5966
                     /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      6137..6725
                     /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 ggggagaaaa gtacattgga
     1501 aactaaaaag atagaatctg aaagtaaagg aatcaaagga ggaaaaaaag tttataaaag
     1561 tttgattact ggaaaagttc gatctaattc agaaatttca ggccaaatga aacagcccct
     1621 tcaagcaaac atgccttcaa tctctcgagg caggacaatg attcatattc caggagttcg
     1681 aaatagctcc tcaagtacaa gtcctgtttc taaaaaaggc ccacccctta agactccagc
     1741 ctccaaaagc cctagtgaag gtcaaacagc caccacttct cctagaggag ccaagccatc
     1801 tgtgaaatca gaattaagcc ctgttgccag gcagacatcc caaataggtg ggtcaagtaa
     1861 agcaccttct agatcaggat ctagagattc gaccccttca agacctgccc agcaaccatt
     1921 aagtagacct atacagtctc ctggccgaaa ctcaatttcc cctggtagaa atggaataag
     1981 tcctcctaac aaattatctc aacttccaag gacatcatcc cctagtactg cttcaactaa
     2041 gtcctcaggt tctggaaaaa tgtcatatac atctccaggt agacagatga gccaacagaa
     2101 ccttaccaaa caaacaggtt tatccaagaa tgccagtagt attccaagaa gtgagtctgc
     2161 ctccaaagga ctaaatcaga tgaataatgg taatggagcc aataaaaagg tagaactttc
     2221 tagaatgtct tcaactaaat caagtggaag tgaatctgat agatcagaaa gacctgtatt
     2281 agtacgccag tcaactttca tcaaagaagc tccaagccca accttaagaa gaaaattgga
     2341 ggaatctgct tcatttgaat ctctttctcc atcatctaga ccagcttctc ccactaggtc
     2401 ccaggcacaa actccagttt taagtccttc ccttcctgat atgtctctat ccacacattc
     2461 gtctgttcag gctggtggat ggcgaaaact cccacctaat ctcagtccca ctatagagta
     2521 taatgatgga agaccagcaa agcgccatga tattgcacgg tctcattctg aaagtccttc
     2581 tagacttcca atcaataggt caggaacctg gaaacgtgag cacagcaaac attcatcatc
     2641 ccttcctcga gtaagcactt ggagaagaac tggaagttca tcttcaattc tttctgcttc
     2701 atcagaatcc agtgaaaaag caaaaagtga ggatgaaaaa catgtgaact ctatttcagg
     2761 aaccaaacaa agtaaagaaa accaagtatc cgcaaaagga acatggagaa aaataaaaga
     2821 aaatgaattt tctcccacaa atagtacttc tcagaccgtt tcctcaggtg ctacaaatgg
     2881 tgctgaatca aagactctaa tttatcaaat ggcacctgct gtttctaaaa cagaggatgt
     2941 ttgggtgaga attgaggact gtcccattaa caatcctaga tctggaagat ctcccacagg
     3001 taatactccc ccggtgattg acagtgtttc agaaaaggca aatccaaaca ttaaagattc
     3061 aaaagataat caggcaaaac aaaatgtggg taatggcagt gttcccatgc gtaccgtggg
     3121 tttggaaaat cgcctgaact cctttattca ggtggatgcc cctgaccaaa aaggaactga
     3181 gataaaacca ggacaaaata atcctgtccc tgtatcagag actaatgaaa gttctatagt
     3241 ggaacgtacc ccattcagtt ctagcagctc aagcaaacac agttcaccta gtgggactgt
     3301 tgctgccaga gtgactcctt ttaattacaa cccaagccct aggaaaagca gcgcagatag
     3361 cacttcagct cggccatctc agatcccaac tccagtgaat aacaacacaa agaagcgaga
     3421 ttccaaaact gacagcacag aatccagtgg aacccaaagt cctaagcgcc attctgggtc
     3481 ttaccttgtg acatctgttt gtacagacta caaagaccat gacggtgatt ataaagatca
     3541 tgacatcgac tacaaggatg acgatgacaa gtaactcgag ataatcaacc tctggattac
     3601 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3661 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3721 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     3781 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa
     3841 atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa tttgtgatgc
     3901 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3961 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagat ctggagaccg
     4021 gtgtggcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4081 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4141 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4201 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4261 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4321 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4441 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4561 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4621 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4681 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4741 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4801 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4861 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4921 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4981 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5041 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5101 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5161 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5221 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5281 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5341 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5401 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5461 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5521 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5581 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5641 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5701 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5761 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5821 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5881 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5941 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6001 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6061 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6781 acatgt
//