pAAVLINKv1-pCALM1-Cre-3'-PCM1-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-PCM1-FLAG               7191 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 7191)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7191
                     /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..3904
                     /codon_start=1
                     /label=PCM1
                     /translation="VK*RNHLSRLDFQCL*KNLQVVTAKIN*IQTEEDASLMKNHWKAL
                     AVCLIQ*IQQQ*LKHSRQEKRLHRPAWHLKIKLPSQKVRRGILLS*KAELKTSGMKVPV
                     CLAHVNLAKVGTDIQPRLKSLFKQKYSAERIMSNWKK**NVIGLQKYLQKLGVIFPCLK
                     LCEILFILK*LH*FLKMNLVHIFLLNSSMSCSY*TQTT*DRGLYMHCRT*YPDIFLRAM
                     KKEKM*SQ*TLVLG*HQTQNLLLVRALLLLMMKLLRRTLKEKPIK*VSKMMLIMLVSCL
                     YHQILSLLQQMI*VTP*FT*IKH*PE*ENMSV*RLRLKVTQI*DAPAGLLRMEMVLVQV
                     LQLII*KKLPLLKIVVHNNL*VKFLPSHVLELILSSWTGKLKQL*KKSFLF*RSTWMKY
                     APRSF*LQ*GAWF*PLPSKMMRAKSL*SSFINNLEVYYRIHWQNLLAEN*KTVEKIFL*
                     RYLKCCSMNWLSLSLCKIWIIIV*LLNRDAKGK*KQLE*YNLVPKRLKGFLKIMAHLLE
                     RLMMKTKTRMKLKQLSRLKHLRCMMVPKM*DLIFLIKRKMKKVKDVQCLLICLKLKLRL
                     *LIMEVEKMKMRMKKWKNLKKALWMSRLPSRLTLKLLKKMNMMNRSYNVTLKRQQKAKM
                     SHWNEKPLVKMTKITVL*NPVTSISWKMSNL*IVLPIRSHLLLLIQLNSLTLCRYVYLK
                     WNP*CLESKKLNLLRKLLKALWLEVLILNLQC**MTMKQNLVI*VKSLMKKIL*KLKIY
                     H*N*QYIQRQI*ERKW*KKNRKTIYLVKYVKCRPKN*LEILRH*KNLKRWEPRV"
     misc_feature    3506..3511
                     /label=SDS
     CDS             3911..3976
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3980..4411
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    4124..4129
                     /label=SDS
     repeat_region   4454..4594
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4454..4583
                     /label=AAV2 ITR
     rep_origin      4669..5124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5406..5510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5511..6371
                     /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      6542..7130
                     /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 ggtgaagtag agaaaccatt
     1501 tatcaagact ggattttcag tgtctgtaga aaaatctaca agtagtaacc gcaaaaatca
     1561 attagataca aacggaagaa gacgccagtt tgatgaagaa tcactggaaa gctttagcag
     1621 tatgcctgat ccagtagatc caacaacagt gactaaaaca ttcaagacaa gaaaagcgtc
     1681 tgcacaggcc agcctggcat ctaaagataa aactcccaag tcaaaaagta agaagaggaa
     1741 ttctactcag ctgaaaagca gagttaaaaa catcaggtat gaaagtgcca gtatgtctag
     1801 cacatgtgaa ccttgcaaaa gtaggaacag acattcagcc cagactgaag agcctgttca
     1861 agcaaaagta ttcagcagaa agaatcatga gcaactggaa aaaataataa aatgtaatag
     1921 gtctacagaa atatcttcag aaactgggag tgatttttcc atgtttgaag ctttgcgaga
     1981 tactatttat tctgaagtag ctacattaat ttctcaaaat gaatctcgtc cacattttct
     2041 tattgaactc ttccatgagc tgcagctact aaacacagac tacttgagac agagggcttt
     2101 atatgcattg caggacatag tatccagaca tatttctgag agccatgaaa aaggagaaaa
     2161 tgtaaagtca gtaaactctg gtacttggat agcatcaaac tcagaactta ctcctagtga
     2221 gagccttgct actactgatg atgaaacttt tgagaagaac tttgaaagag aaacccataa
     2281 aataagtgag caaaatgatg ctgataatgc tagtgtcctg tctgtatcat caaattttga
     2341 gccttttgca acagatgatc taggtaacac cgtgattcac ttagatcaag cattagccag
     2401 aatgagagaa tatgagcgta tgaagactga ggctgaaagt aactcaaata tgagatgcac
     2461 ctgcaggatt attgaggatg gagatggtgc tggtgcaggt actacagtta ataatttaga
     2521 agaaactccc gttattgaaa atcgtagttc acaacaacct gtaagtgaag tttctaccat
     2581 cccatgtcct agaattgata ctcagcagct ggaccggcaa attaaagcaa ttatgaaaga
     2641 agtcattcct tttttgaagg agcacatgga tgaagtatgc tcctcgcagc ttctaacttc
     2701 agtaaggcgc atggttttga cccttaccca gcaaaatgat gagagcaaag agtttgtaaa
     2761 gttctttcat aaacaacttg gaagtatatt acaggattca ctggcaaaat ttgctggcag
     2821 aaaactgaaa gactgtggag aagatcttct tgtagagata tctgaagtgt tgttcaatga
     2881 attggctttc tttaagctta tgcaagattt ggataataat agtataactg ttaaacagag
     2941 atgcaaaagg aaaatagaag caactggagt gatacaatct tgtgccaaag aggctaaaag
     3001 gattcttgaa gatcatggct cacctgctgg agagattgat gatgaagaca aagacaagga
     3061 tgaaactgaa acagttaagc agactcaaac atctgaggtg tatgatggtc ccaaaaatgt
     3121 aagatctgat atttctgatc aagaggaaga tgaagaaagt gaaggatgtc cagtgtctat
     3181 taatttgtct aaagctgaaa ctcaggcttt aactaattat ggaagtggag aagatgaaaa
     3241 tgaggatgaa gaaatggaag aatttgaaga aggccctgtg gatgtccaga cttccctcca
     3301 ggctaacact gaagctactg aagaaaatga acatgatgaa caggtcctac aacgtgactt
     3361 taaaaagaca gcagaaagca aaaatgtccc attggaacga gaagccacta gtaaaaatga
     3421 ccaaaataac tgtcctgtga aaccctgtta cctcaatatc ttggaagatg agcaaccttt
     3481 aaatagtgct gcccataagg agtcacctcc tactgttgat tcaactcaac agcctaaccc
     3541 tttgccgtta cgtttacctg aaatggaacc cttagtgcct agagtcaaag aagttaaatc
     3601 tgctcaggaa actcctgaaa gctctctggc tggaagtcct gatactgaat ctccagtgtt
     3661 agtgaatgac tatgaagcag aatctggtaa tataagtcaa aagtctgatg aagaagattt
     3721 tgtaaaagtt gaagatttac cactgaaact gacaatatat tcagaggcag atctaagaaa
     3781 gaaaatggta gaagaagaac agaaaaacca tttatctggt gaaatatgtg aaatgcagac
     3841 cgaagaatta gctggaaatt ctgagacact aaaagaacct gaaacggtgg gagcccagag
     3901 tatatgtaca gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3961 ggatgacgat gacaagtaac tcgagataat caacctctgg attacaaaat ttgtgaaaga
     4021 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4081 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4141 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4201 gctcggctgt tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt
     4261 gctttatttg taaccatcta gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4321 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4381 ggttcagggg gagatgtggg aggtttttta aagatctgga gaccggtgtg gcacgtgcgg
     4441 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4501 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4561 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4621 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4681 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4741 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4801 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4861 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4921 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4981 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     5041 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5101 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5161 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5221 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5281 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5401 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5461 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5761 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6061 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6121 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//