pAAVLINKv1-pCALM1-Cre-3'-TNS2-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-TNS2-FLAG                6849 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 6849)
  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..6849
                     /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..3562
                     /codon_start=1
                     /label=TNS2
                     /translation="GGQVRLASLSCTQCGLGTRCLCSCLPVGITMPRCLTTAA*SHPRQ
                     ARKGTRAAPTPCAPKAGMGIQGTLPW*HTAMEEQFPVTAQHMAVCLIAVALQERAEGIP
                     ALVPTPHGLAPFPRAARPIHNLGS*ATRSLRRREGTGTHCLGTWPQQDLWHLQSRWSRC
                     PGGRAPVGTAHCLGLPEMPHAVLRQSCLVPPRPCTPAVQSRARKAPGDRTPGPPPQRPL
                     RD*VLARPCPLFPRQAPERPLSCRREVGLSLWPLAQSLRPSLPARPVTGLRKGVQGATQ
                     MAPVLGALCPPHFLASATPPGKALEAPPTAQMGLPSLLCLPRCPGLWPAQSRLRAHLHL
                     LSPWLPPMTPMALASPHFLRNATCPGRGNSQDPGAQSRHHRQPEASVTMSPSHLCSQIM
                     SPKPQSLLHKRAKAMSSLSRIHPSSGTSHTCPVTKPLPC*RTRTLGPS*SGTVIHSKEL
                     MGWPSRWPHRHPVPSPGKGTPWNSWSAISSSRLGPKG*RSRAAPVSPTLAACPPWSPST
                     PSPPSPCPAACAFPAKILWKRPQRLQCPPT*AQRQTSCVRVLPAACST*PQWRQSH*RA
                     PKLWPGPALQL*AVAPARHQLLSTSRCQPRALH*RTTKGSSSFAAIIQ*TASPSPALTL
                     KTGDGPTQTGPPPRSLVSWPRSREAPGRMCVTSLQSLTQISLLAPLSPSSPKFYWARE"
     misc_feature    1887..1892
                     /label=SDS
     misc_feature    2517..2522
                     /label=SDS
     misc_feature    2696..2701
                     /label=SDS
     misc_feature    3139..3144
                     /label=SDS
     CDS             3569..3634
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3638..4069
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3782..3787
                     /label=SDS
     repeat_region   4112..4252
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4112..4241
                     /label=AAV2 ITR
     rep_origin      4327..4782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5064..5168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5169..6029
                     /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      6200..6788
                     /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 gggtgggcaa gtgaggctgg
     1501 caagcctctc ctgcacccag tgcggcctgg gcacccgctg cctctgctct tgcctgcctg
     1561 tgggcatcac catgccccga tgcctgacta cagctgcctg aagccaccca aggcaggcga
     1621 ggaagggcac gagggctgct cctacaccat gtgccccgaa ggcaggtatg ggcatccagg
     1681 gtaccctgcc ctggtgacat acagctatgg aggagcagtt cccagttact gcccagcata
     1741 tggccgtgtg cctcatagct gtggctctcc aggagagggc agagggtatc ccagccctgg
     1801 tgcccactcc ccacgggctg gctccatttc cccgggcagc ccgccctatc cacaatctag
     1861 gaagctgagc tacgagatcc ctacggagga gggaggggac aggtacccat tgcctgggca
     1921 cctggcctca gcaggacctt tggcatctgc agagtcgctg gagccggtgt cctggaggga
     1981 gggccccagt gggcacagca cactgcctcg gtctccccga gatgccccat gcagtgcttc
     2041 gtcagagttg tctggtccct ccacgcccct gcacaccagc agtccagtcc agggcaagga
     2101 aagcacccgg cgacaggaca ccaggtcccc cacctcagcg cccactcaga gactgagtcc
     2161 tggcgaggcc ttgccccctg tttcccaggc aggcaccgga aaggcccctg agctgccgtc
     2221 gggaagtggg cctgagcctc tggcccctag cccagtctct ccgaccttcc ctcccagctc
     2281 gcccagtgac tggcctcagg aaaggagtcc agggggccac tcagatggcg ccagtcctcg
     2341 gagccctgtg cccaccacac ttcctggcct ccgccacgcc ccctggcaag gccctcgagg
     2401 cccccccgac agcccagatg ggtctcccct cactcctgtg ccttcccaga tgccctggct
     2461 tgtggccagc ccagagccgc ctcagagctc acctacacct gctttccccc tggctgcctc
     2521 ctatgacacc aatggcctta gccagccccc acttcctgag aaacgccacc tgcccgggcc
     2581 ggggcaacag ccaggaccct ggggcccaga gcaggcatca tcgccagcca gaggcatcag
     2641 tcaccatgtc accttcgcac ctctgctctc agataatgtc ccccaaaccc cagagcctcc
     2701 tacacaagag agccaaagca atgtcaagtt tgtccaggat acatccaagt tctggtacaa
     2761 gccacacctg tcccgtgacc aagccattgc cctgctgaag gacaaggacc ctggggcctt
     2821 cctgatcagg gacagtcatt cattccaagg agcttatggg ctggccctca aggtggccac
     2881 accgccaccc agtgcccagc cctggaaagg ggaccccgtg gaacagctgg tccgccattt
     2941 cctcatcgag actgggccca aaggggtgaa gatcaagggc tgccccagtg agccctactt
     3001 tggcagcctg tccgccttgg tctcccagca ctccatctcc cccatctccc tgccctgctg
     3061 cctgcgcatt cccagcaaag atcctctgga agagacccca gaggctccag tgcccaccaa
     3121 catgagcaca gcggcagacc tcctgcgtca gggtgctgcc tgcagcgtgc tctacttgac
     3181 ctcagtggag acagagtcac tgacgggccc ccaagctgtg gcccgggcca gctctgcagc
     3241 tctgagctgt agcccccgcc cgacaccagc tgttgtccac ttcaaggtgt cagcccaggg
     3301 cattacactg acggacaacc aaaggaagct cttctttcgc cgccattatc cagtgaacag
     3361 catcaccttc tccagcactg accctcaaga ccggagatgg accaacccag acgggaccac
     3421 ctccaagatc tttggtttcg tggccaagaa gccgggaagc ccctgggaga atgtgtgtca
     3481 cctctttgca gagcttgacc cagatcagcc tgctggcgcc attgtcacct tcatcaccaa
     3541 agttctactg ggccagagaa aatgtacaga ctacaaagac catgacggtg attataaaga
     3601 tcatgacatc gactacaagg atgacgatga caagtaactc gagataatca acctctggat
     3661 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3721 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3781 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     3841 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gtttatttgt
     3901 gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg aaatttgtga
     3961 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4021 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gatctggaga
     4081 ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4141 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4201 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4261 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4321 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4381 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4441 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4501 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4561 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4621 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4681 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4741 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4801 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4861 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4921 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4981 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5161 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5221 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5281 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5341 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5401 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5461 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5521 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5581 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5641 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5701 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5761 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5821 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5881 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5941 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6001 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6061 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6841 ctcacatgt
//