pAAVLINKv1-pCALM1-Cre-3'-MYLK-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-MYLK-FLAG                6766 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 6766)
  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..6766
                     /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..3479
                     /codon_start=1
                     /label=MYLK
                     /translation="VRAGESVELFGKVTGTQPITCTWMKFRKQIQESEHMKVENSENGS
                     KLTILAARQEHCGCYTLLVENKLGSRQAQVNLTVVDKPDPPAGTPCASDIRSSSLTLSW
                     YGSSYDGGSAVQSYSIEIWDSANKTWKELATCRSTSFNVQDLLPDHEYKFRVRAINVYG
                     TSEPSQESELTTVGEKPEEPKDEVEVSDDDEKEPEVDYRTVTINTEQKVSDFYDIEERL
                     GSGKFGQVFRLVEKKTRKVWAGKFFKAYSAKEKENIRQEISIMNCLHHPKLVQCVDAFE
                     EKANIVMVLEIVSGGELFERIIDEDFELTERECIKYMRQISEGVEYIHKQGIVHLDLKP
                     ENIMCVNKTGTRIKLIDFGLARRLENAGSLKVLFGTPEFVAPEVINYEPIGYATDMWSI
                     GVICYILVSGLSPFMGDNDNETLANVTSATWDFDDEAFDEISDDAKDFISNLLKKDMKN
                     RLDCTQCLQHPWLMKDTKNMEAKKLSKDRMKKYMARRKWQKTGNAVRAIGRLSSMAMIS
                     GLSGRKSSTGSPTSPLNAEKLESEEDVSQAFLEAVAEEKPHVKPYFSKTIRDLEVVEGS
                     AARFDCKIEGYPDPEVVWFKDDQSIRESRHFQIDYDEDGNCSLIISDVCGDDDAKYTCK
                     AVNSLGEATCTAELIVETMEEGEGEGEEEEE"
     misc_feature    2363..2368
                     /label=SDS
     misc_feature    2565..2570
                     /label=SDS
     CDS             3486..3551
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3555..3986
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3699..3704
                     /label=SDS
     repeat_region   4029..4169
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4029..4158
                     /label=AAV2 ITR
     rep_origin      4244..4699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4981..5085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5086..5946
                     /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      6117..6705
                     /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 ggtacgcgca ggagagtcag
     1501 tggagctgtt tggcaaagtg acaggcactc agcccatcac ctgtacctgg atgaagttcc
     1561 gaaagcagat ccaggaaagc gagcacatga aggtggagaa cagcgagaat ggcagcaagc
     1621 tcaccatcct ggccgcgcgc caggagcact gcggctgcta cacactgctg gtggagaaca
     1681 agctgggcag caggcaggcc caggtcaacc tcactgtcgt ggataagcca gaccccccag
     1741 ctggcacacc ttgtgcctct gacattcgga gctcctcact gaccctgtcc tggtatggct
     1801 cctcatatga tgggggcagt gctgtacagt cctacagcat cgagatctgg gactcagcca
     1861 acaagacgtg gaaggaacta gccacatgcc gcagcacctc tttcaacgtc caggacctgc
     1921 tgcctgacca cgaatataag ttccgtgtac gtgcaatcaa cgtgtatgga accagtgagc
     1981 caagccagga gtctgaactc acaacggtag gagagaaacc tgaagagccg aaggatgaag
     2041 tggaggtgtc agatgatgat gagaaggagc ccgaggttga ttaccggaca gtgacaatca
     2101 atactgaaca aaaagtatct gacttctacg acattgagga gagattagga tctgggaaat
     2161 ttggacaggt ctttcgactt gtagaaaaga aaactcgaaa agtctgggca gggaagttct
     2221 tcaaggcata ttcagcaaaa gagaaagaga atatccggca ggagattagc atcatgaact
     2281 gcctccacca ccctaagctg gtccagtgtg tggatgcctt tgaagaaaag gccaacatcg
     2341 tcatggtcct ggagatcgtg tcaggagggg agctgtttga gcgcatcatt gacgaggact
     2401 ttgagctgac ggagcgtgag tgcatcaagt acatgcggca gatctcggag ggagtggagt
     2461 acatccacaa gcagggcatc gtgcacctgg acctcaagcc ggagaacatc atgtgtgtca
     2521 acaagacggg caccaggatc aagctcatcg actttggtct ggccaggagg ctggagaatg
     2581 cggggtctct gaaggtcctc tttggcaccc cagaatttgt ggctcctgaa gtgatcaact
     2641 atgagcccat cggctacgcc acagacatgt ggagcatcgg ggtcatctgc tacatcctag
     2701 tcagtggcct ttcccccttc atgggagaca acgataacga aaccttggcc aacgttacct
     2761 cagccacctg ggacttcgac gacgaggcat tcgatgagat ctccgacgat gccaaggatt
     2821 tcatcagcaa tctgctgaag aaagatatga aaaaccgcct ggactgcacg cagtgccttc
     2881 agcatccatg gctaatgaaa gataccaaga acatggaggc caagaaactc tccaaggacc
     2941 ggatgaagaa gtacatggca agaaggaaat ggcagaaaac gggcaatgct gtgagagcca
     3001 ttggaagact gtcctctatg gcaatgatct cagggctcag tggcaggaaa tcctcaacag
     3061 ggtcaccaac cagcccgctc aatgcagaaa aactagaatc tgaagaagat gtgtcccaag
     3121 ctttccttga ggctgttgct gaggaaaagc ctcatgtaaa accctatttc tctaagacca
     3181 ttcgcgattt agaagttgtg gagggaagtg ctgctagatt tgactgcaag attgaaggat
     3241 acccagaccc cgaggttgtc tggttcaaag atgaccagtc aatcagggag tcccgccact
     3301 tccagataga ctacgatgag gacgggaact gctctttaat tattagtgat gtttgcgggg
     3361 atgacgatgc caagtacacc tgcaaggctg tcaacagtct tggagaagcc acctgcacag
     3421 cagagctcat tgtggaaacg atggaggaag gtgaagggga aggggaagag gaagaagagt
     3481 gtacagacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3541 acgatgacaa gtaactcgag ataatcaacc tctggattac aaaatttgtg aaagattgac
     3601 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3661 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3721 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3781 gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt
     3841 atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3901 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3961 agggggagat gtgggaggtt ttttaaagat ctggagaccg gtgtggcacg tgcggaccga
     4021 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4081 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4141 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4201 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4261 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4321 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4381 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4441 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4501 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4561 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4621 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4681 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4741 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4801 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4861 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4921 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4981 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5041 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5101 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5161 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5221 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5281 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5341 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5401 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5461 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5521 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5581 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5641 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5701 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5761 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5821 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5881 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5941 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6001 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6061 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6241 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6421 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6481 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6541 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6601 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6661 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6721 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//