pAAVLINKv1-pCALM1-Cre-3'-COL5A1-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-COL5A1-FLAG               6871 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 6871)
  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..6871
                     /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..3584
                     /codon_start=1
                     /label=COL5A1
                     /translation="GPVGLPGPAGPVGPPGEDGDKGEIGEPGQKGSKGDKGEQGPPGPT
                     GPQGPIGQPGPSGADGEPGPRGQQGLFGQKGDEGPRGFPGPPGPVGLQGLPGPPGEKGE
                     TGDVGQMGPPGPPGPRGPSGAPGADGPQGPPGGIGNPGAVGEKGEPGEAGEPGLPGEGG
                     PPGPKGERGEKGESGPSGAAGPPGPKGPPGDDGPKGSPGPVGFPGDPGPPGEPGPAGQD
                     GPPGDKGDDGEPGQTGSPGPTGEPGPSGPPGKRGPPGPAGPEGRQGEKGAKGEAGLEGP
                     PGKTGPIGPQGAPGKPGPDGLRGIPGPVGEQGLPGSPGPDGPPGPMGPPGLPGLKGDSG
                     PKGEKGHPGLIGLIGPPGEQGEKGDRGLPGPQGSSGPKGEQGITGPSGPIGPPGPPGLP
                     GPPGPKGAKGSSGPTGPKGEAGHPGPPGPPGPPGEVIQPLPIQASRTRRNIDASQLLDD
                     GNGENYVDYADGMEEIFGSLNSLKLEIEQMKRPLGTQQNPARTCKDLQLCHPDFPDGEY
                     WVDPNQGCSRDSFKVYCNFTAGGSTCVFPDKKSEGARITSWPKENPGSWFSEFKRGKLL
                     SYVDAEGNPVGVVQMTFLRLLSASAHQNVTYHCYQSVAWQDAATGSYDKALRFLGSNDE
                     EMSYDNNPYIRALVDGCATKKGYQKTVLEIDTPKVEQVPIVDIMFNDFGEASQKFGFEV
                     GPACFMG"
     misc_feature    1602..1607
                     /label=SDS
     misc_feature    2322..2327
                     /label=SDS
     misc_feature    2655..2660
                     /label=SDS
     CDS             3591..3656
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3660..4091
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3804..3809
                     /label=SDS
     repeat_region   4134..4274
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4134..4263
                     /label=AAV2 ITR
     rep_origin      4349..4804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5086..5190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5191..6051
                     /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      6222..6810
                     /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 ggggcctgtg gggctcccgg
     1501 gtccagctgg ccctgtgggt ccccctggag aagacggaga taagggagag atcggggagc
     1561 cggggcagaa aggaagcaag ggggacaaag gagaacaggg tcctcctggg cctacaggtc
     1621 ctcaaggccc catcggacag ccaggcccct ctggagctga cggcgagccg gggcctcggg
     1681 gccagcaggg ccttttcggg cagaaaggtg atgaaggtcc cagaggcttt cctggacccc
     1741 ctgggccagt ggggctgcag ggtttgccag gacctccagg cgagaagggt gagacaggag
     1801 acgtgggcca gatgggcccc ccgggtcccc ctggcccccg aggaccctcc ggagctccag
     1861 gtgctgatgg cccacaaggt cccccaggtg gaataggaaa ccctggtgca gtgggagaga
     1921 agggcgagcc tggcgaagca ggtgagcctg gccttccggg agaaggcggc cccccgggac
     1981 ccaaaggaga aaggggagag aagggcgagt caggcccttc aggtgctgcc ggaccccctg
     2041 gacccaaagg ccctcccgga gatgatggtc ccaaaggcag ccctggccca gtgggttttc
     2101 ctggagatcc tggccccccc ggagagcctg gccccgcggg tcaagatggt ccccctggtg
     2161 acaaaggaga tgatggtgaa cccgggcaga cgggatcccc cggccctact ggtgaaccag
     2221 gtccatcggg gcctccagga aaaaggggtc ccccaggccc cgcaggcccc gaaggcagac
     2281 agggagagaa aggggccaag ggagaagccg gcttggaagg ccctcctggg aagactggcc
     2341 ccatcggccc ccagggggcc cctgggaagc ccggaccgga tggccttcga gggatccctg
     2401 gccctgtggg agaacaaggt ctcccaggat ccccaggccc ggacggtccc cccggcccca
     2461 tgggtccccc aggacttccc ggcctcaaag gagattctgg tcccaaaggt gaaaagggtc
     2521 atccaggcct gatcgggctc atcggtcctc ccggtgaaca gggtgagaag ggcgaccgtg
     2581 gtctccctgg cccccagggc tcctccggtc ctaagggaga acagggtatc actggtcctt
     2641 ctggcccgat tgggcctcct gggccccctg gcctgccggg tccgcctggt ccaaaaggtg
     2701 ctaagggctc ctcgggtcca actggcccga agggtgaggc aggccaccca ggacccccag
     2761 gccccccggg ccccccggga gaggtcatcc agcccctgcc aatccaggca tccaggacgc
     2821 ggcggaacat cgacgccagc cagctgctgg acgacgggaa tggcgagaac tacgtggact
     2881 acgcggacgg catggaagag atcttcggct ctctcaactc tctgaagctg gagattgagc
     2941 agatgaaacg gcccctgggc acgcagcaga accccgcccg cacctgcaag gacctgcagc
     3001 tctgccaccc cgacttccca gatggtgaat actgggtcga tcctaaccaa ggatgctcca
     3061 gggattcctt caaggtttac tgcaacttca cagccggggg gtcgacatgc gtcttccctg
     3121 acaagaagtc cgaaggggcc agaatcactt cttggcccaa agaaaacccg ggctcctggt
     3181 tcagtgaatt caagcgtggg aaactgctct cctatgtgga cgccgagggc aaccctgtgg
     3241 gtgtggtaca gatgaccttc ctgcggctgc tgagcgcctc tgcccaccag aacgtcacct
     3301 accactgcta ccagtcagtg gcctggcagg acgcagccac gggcagctac gacaaggccc
     3361 tccgcttcct gggctccaac gacgaggaga tgtcctatga caacaacccc tacatccgcg
     3421 ccctggtgga cggctgtgct accaagaaag gctaccagaa gacggttctg gagatcgaca
     3481 cccccaaagt ggagcaggtg cccatcgtgg acatcatgtt caatgacttc ggtgaagcgt
     3541 cacagaaatt tggatttgaa gtggggccgg cttgcttcat gggctgtaca gactacaaag
     3601 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtaac
     3661 tcgagataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3721 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3781 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     3841 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3901 caattccgtg gtgtttattt gtgaaatttg tgatgctatt gctttatttg taaccatcta
     3961 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4021 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     4081 aggtttttta aagatctgga gaccggtgtg gcacgtgcgg accgagcggc cgcaggaacc
     4141 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4201 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4261 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4321 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4381 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4441 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4501 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4561 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4621 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4681 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4741 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4801 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4861 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4921 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4981 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5041 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5101 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5161 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5221 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5281 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5341 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5401 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5461 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5521 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5581 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5641 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5701 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5761 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5821 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5881 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5941 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6001 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6061 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6121 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6841 ctggcctttt gctggccttt tgctcacatg t
//