pAAVLINKv1-pCALM1-Cre-3'-BLM-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-BLM-FLAG             6348 bp DNA     circular SYN 18-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 1, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6348
                     /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..3061
                     /codon_start=1
                     /label=BLM
                     /translation="G*FTASPGENVTPWLTRYREMGSLLLLTMLASVILPEMKCSRSGL
                     IRMAVRLSVLQLHLEWGLTNRTCDL*FMHLSLNLWRVTTKNLAELEEMGKYLTACFSIP
                     IMM*PD*KDL**WKKMETIIQEKLTSIICIAWYITVKI*RNAGEYSFWPTLVKMDLILI
                     FVRNTQMFLVIIAVKQRIIKQEM*LTM*KVL*DLFKNIVHHKE*EI*NM*VLLEDLL*I
                     CWSTFSWGVRVQKSSQVYLEKDLLIHDTMPKDFLKS*YLTRFWMKTYISMPMTRRSLM*
                     CSEIKPKLY*MAI*R*TLWKQKIPAV*KNKKR**QKCLRGKRWLKNVLENLQKSANLWG
                     KFLVSITSIFLIPSLSRSLQNLYLLILRFCFKLMVLLKTNWKNMVRK*FQYYRNTLNGH
                     RQLKTVPQG*ACPAAEAPEEVPLRSSTRKYPYLPTTLQVKPEMKGRGKRCQPPKGLRGE
                     KLLPVVPRQRGGLPHVERYLPKRNPPASLDPVQPHILLKRHQEPIANWGLWLHRSL*ID
                     RFLSLHMHS"
     CDS             3068..3133
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3137..3568
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3281..3286
                     /label=SDS
     repeat_region   3611..3751
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    3611..3740
                     /label=AAV2 ITR
     rep_origin      3826..4281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4563..4667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4668..5528
                     /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      5699..6287
                     /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 gggataattt actgcctctc
     1501 caggcgagaa tgtgacacca tggctgacac gttacagaga gatgggctcg ctgctcttgc
     1561 ttaccatgct ggcctcagtg attctgccag agatgaagtg cagcagaagt ggattaatca
     1621 ggatggctgt caggttatct gtgctacaat tgcatttgga atggggattg acaaaccgga
     1681 cgtgcgattt gtgattcatg catctctccc taaatctgtg gagggttact accaagaatc
     1741 tggcagagct ggaagagatg gggaaatatc tcactgcctg cttttctata cctatcatga
     1801 tgtgaccaga ctgaaaagac ttataatgat ggaaaaagat ggaaaccatc atacaagaga
     1861 aactcacttc aataatttgt atagcatggt acattactgt gaaaatataa cggaatgcag
     1921 gagaatacag cttttggcct actttggtga aaatggattt aatcctgatt tttgtaagaa
     1981 acacccagat gtttcttgtg ataattgctg taaaacaaag gattataaaa caagagatgt
     2041 gactgacgat gtgaaaagta ttgtaagatt tgttcaagaa catagttcat cacaaggaat
     2101 gagaaatata aaacatgtag gtccttctgg aagatttact atgaatatgc tggtcgacat
     2161 tttcttgggg agtaagagtg caaaaatcca gtcaggtata tttggaaaag gatctgctta
     2221 ttcacgacac aatgccgaaa gactttttaa aaagctgata cttgacaaga ttttggatga
     2281 agacttatat atcaatgcca atgaccaggc gatcgcttat gtgatgctcg gaaataaagc
     2341 ccaaactgta ctaaatggca atttaaaggt agactttatg gaaacagaaa attccagcag
     2401 tgtgaaaaaa caaaaagcgt tagtagcaaa agtgtctcag agggaagaga tggttaaaaa
     2461 atgtcttgga gaacttacag aagtctgcaa atctctgggg aaagtttttg gtgtccatta
     2521 cttcaatatt tttaataccg tcactctcaa gaagcttgca gaatctttat cttctgatcc
     2581 tgaggttttg cttcaaattg atggtgttac tgaagacaaa ctggaaaaat atggtgcgga
     2641 agtgatttca gtattacaga aatactctga atggacatcg ccagctgaag acagttcccc
     2701 agggataagc ctgtccagca gcagaggccc cggaagaagt gccgctgagg agctcgacga
     2761 ggaaataccc gtatcttccc actactttgc aagtaaaacc agaaatgaaa ggaagaggaa
     2821 aaagatgcca gcctcccaaa ggtctaagag gagaaaaact gcttccagtg gttccaaggc
     2881 aaaggggggg tctgccacat gtagaaagat atcttccaaa acgaaatcct ccagcatcat
     2941 tggatccagt tcagcctcac atacttctca agcgacatca ggagccaata gcaaattggg
     3001 gattatggct ccaccgaagc ctataaatag accgtttctt aagccttcat atgcattctc
     3061 atgtacagac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3121 tgacgatgac aagtaactcg agataatcaa cctctggatt acaaaatttg tgaaagattg
     3181 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3241 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3301 ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3361 cggctgttgg gcactgacaa ttccgtggtg tttatttgtg aaatttgtga tgctattgct
     3421 ttatttgtaa ccatctagct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3481 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3541 tcagggggag atgtgggagg ttttttaaag atctggagac cggtgtggca cgtgcggacc
     3601 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3661 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3721 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3781 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3841 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3901 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3961 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4021 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     4081 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4141 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     4201 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4261 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4321 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4381 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4441 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4501 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5521 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5581 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5821 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//