pAAVLINKv1-pCALM1-Cre-3'-AGL-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-AGL-FLAG               6885 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 6885)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6885
                     /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..3598
                     /codon_start=1
                     /label=AGL
                     /translation="GPMNIFKK*NLKTCLQEVLLYSELVLIHMHKSLLEFFEII*HNSV
                     LTLNLAA*LLTMQILY*KFLLLLLPPD*LWLS*IRSFTDVNQKKRKMVEGAMTYQTGQP
                     LNMQVFKV*CLYWQK*DQRMTWGILFVII*DLEIG*LTMSVTGLFHDQELLLKLVNGCR
                     LCSST*SRSHVTLSHVTLMLY*LVHIPLFWIQHGSRCQALFRMVQPL*NTFHWVQFNCV
                     E*ENSLPCQFFHLP*WMYLIG*MRSQKKRSNVVFL*LQAYLIFLLVFSAAGEGILLLHL
                     EVYC*LLDAM*KPGILF*HLRVP*GMVSFLIYWVKEFMPDTIVGMLCGGGCSVSRITVK
                     WFQMV*TFSSAQFPECILQMILLLCLLAHWISHCLKSYRKQCKNTCRAYSSEKGMLVPR
                     *IET*RTKVLI*LQELMKKQDLFMEEIVSIVAHGWIKWEKVTELETEESQPHQEMGLLW
                     KLWA*VNLLFAGCWNYPKKIFSLIMKSQ*KDMERL*RSHMMSGTEKYKTTLKSYFMFPK
                     TLQI*MKSIQIWFTNVAYTKIVMELQVLGVTISSGLILP*QWLWPLSSLLQKKHGKLWR
                     LQKKNCLVPLA*KL*IQMIWFTVEFMTMH*TMTTTILLKVSIITKDLSGCGLLGIFFVQ
                     NYIFPD*WARRLLQRL*FWLKMFFPDIMFILRDPLGKDFQN*PMRMPSTVLSAVKHKPG
                     QLLLFLRHFMI"
     CDS             3605..3670
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3674..4105
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3818..3823
                     /label=SDS
     repeat_region   4148..4288
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4148..4277
                     /label=AAV2 ITR
     rep_origin      4363..4818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5100..5204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5205..6065
                     /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      6236..6824
                     /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 gggcccaatg aatatattca
     1501 agaaatagaa tttgaaaact tgtctccagg aagtgttatt atattcagag ttagtcttga
     1561 tccacatgca caagtcgctg ttggaattct tcgaaatcat ctgacacaat tcagtcctca
     1621 ctttaaatct ggcagcctag ctgttgacaa tgcagatcct atattaaaaa ttccttttgc
     1681 ttctcttgcc tccagattaa ctttggctga gctaaatcag atcctttacc gatgtgaatc
     1741 agaagaaaag gaagatggtg gagggtgcta tgacatacca aactggtcag cccttaaata
     1801 tgcaggtctt caaggtttaa tgtctgtatt ggcagaaata agaccaaaga atgacttggg
     1861 gcatcctttt tgtaataatt tgagatctgg agattggatg attgactatg tcagtaaccg
     1921 gcttatttca cgatcaggaa ctattgctga agttggtaaa tggttgcagg ctatgttctt
     1981 ctacctgaag cagatcccac gttaccttat cccatgttac tttgatgcta tattaattgg
     2041 tgcatatacc actcttctgg atacagcatg gaagcagatg tcaagctttg ttcagaatgg
     2101 ttcaaccttt gtgaaacacc tttcattggg ttcagttcaa ctgtgtggag taggaaaatt
     2161 cccttccctg ccaattcttt cacctgccct aatggatgta ccttataggt taaatgagat
     2221 cacaaaagaa aaggagcaat gttgtgtttc tctagctgca ggcttacctc atttttcttc
     2281 tggtattttc cgctgctggg gaagggatac ttttattgca cttagaggta tactgctgat
     2341 tactggacgc tatgtagaag ccaggaatat tattttagca tttgcgggta ccctgaggca
     2401 tggtctcatt cctaatctac tgggtgaagg aatttatgcc agatacaatt gtcgggatgc
     2461 tgtgtggtgg tggctgcagt gtatccagga ttactgtaaa atggttccaa atggtctaga
     2521 cattctcaag tgcccagttt ccagaatgta tcctacagat gattctgctc ctttgcctgc
     2581 tggcacactg gatcagccat tgtttgaagt catacaggaa gcaatgcaaa aacacatgca
     2641 gggcatacag ttccgagaaa ggaatgctgg tccccagata gatcgaaaca tgaaggacga
     2701 aggttttaat ataactgcag gagttgatga agaaacagga tttgtttatg gaggaaatcg
     2761 tttcaattgt ggcacatgga tggataaaat gggagaaagt gacagagcta gaaacagagg
     2821 aatcccagcc acaccaagag atgggtctgc tgtggaaatt gtgggcctga gtaaatctgc
     2881 tgttcgctgg ttgctggaat tatccaaaaa aaatattttc ccttatcatg aagtcacagt
     2941 aaaaagacat ggaaaggcta taaaggtctc atatgatgag tggaacagaa aaatacaaga
     3001 caactttgaa aagctatttc atgtttccga agacccttca gatttaaatg aaaagcatcc
     3061 aaatctggtt cacaaacgtg gcatatacaa agatagttat ggagcttcaa gtccttggtg
     3121 tgactatcag ctcaggccta attttaccat agcaatggtt gtggcccctg agctctttac
     3181 tacagaaaaa gcatggaaag ctttggagat tgcagaaaaa aaattgcttg gtccccttgg
     3241 catgaaaact ttagatccag atgatatggt ttactgtgga atttatgaca atgcattaga
     3301 caatgacaac tacaatcttg ctaaaggttt caattatcac caaggacctg agtggctgtg
     3361 gcctattggg tattttcttc gtgcaaaatt atatttttcc agattgatgg gcccggagac
     3421 tactgcaaag actatagttt tggttaaaaa tgttctttcc cgacattatg ttcatcttga
     3481 gagatcccct tggaaaggac ttccagaact gaccaatgag aatgcccagt actgtccttt
     3541 cagctgtgaa acacaagcct ggtcaattgc tactattctt gagacacttt atgatttatg
     3601 tacagactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3661 cgatgacaag taactcgaga taatcaacct ctggattaca aaatttgtga aagattgact
     3721 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3781 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     3841 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3901 ctgttgggca ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta
     3961 tttgtaacca tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4021 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4081 gggggagatg tgggaggttt tttaaagatc tggagaccgg tgtggcacgt gcggaccgag
     4141 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4201 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4261 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4321 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     4441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4561 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4681 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4741 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4801 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4861 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4921 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4981 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5041 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5101 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5161 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5221 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5281 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5341 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5401 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5461 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5521 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5581 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5641 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5701 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5761 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5821 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5881 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5941 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6001 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6061 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6121 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6181 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6241 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6301 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6361 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6421 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6481 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6541 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6601 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6661 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6721 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6781 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6841 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//