pAAVLINKv1-pCALM1-Cre-3'-TRIP12-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-TRIP12-FLAG            7345 bp DNA     circular SYN 10-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 9, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7345
                     /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"
                     /note="color: #ffe4c4"
     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..4058
                     /codon_start=1
                     /gene="TRIP12"
                     /gene_synonym="MRD49; TRIP-12; TRIPC; ULF"
                     /product="E3 ubiquitin-protein ligase TRIP12 isoform h"
                     /EC_number="2.3.2.26"
                     /label=TRIP12
                     /note="isoform h is encoded by transcript variant 
                     13;thyroid receptor interacting protein 12; probable E3 
                     ubiquitin-protein ligase TRIP12; TR-interacting protein 12;
                     E3 ubiquitin-protein ligase for Arf; HECT-type E3 ubiquitin
                     transferase TRIP12"
                     /db_xref="GI:1135449777"
                     /db_xref="CCDS:CCDS92966.1"
                     /db_xref="GeneID:9320"
                     /db_xref="HGNC:HGNC:12306"
                     /db_xref="MIM:604506"
                     /protein_id="NP_001335252.1"
                     /translation="VDGGAECLVEIRSIVSESDVSSFEIQHSGFVKQLLLYLTSKSEKD
                     AVSREIRLKRFLHVFFSSPLPGEEPIGRVEPVGNAPLLALVHKMNNCLSQMEQFPVKVH
                     DFPSGNGTGGSFSLNRGSQALKFFNTHQLKCQLQRHPDCANVKQWKGGPVKIDPLALVQ
                     AIERYLVVRGYGRVREDDEDSDDDGSDEEIDESLAAQFLNSGNVRHRLQFYIGEHLLPY
                     NMTVYQAVRQFSIQAEDERESTDDESNPLGRAGIWTKTHTIWYKPVREDEESNKDCVGG
                     KRGRAQTAPTKTSPRNAKKHDELWHDGVCPSVSNPLEVYLIPTPPENITFEDPSLDVIL
                     LLRVLHAISRYWYYLYDNAMCKEIIPTSEFINSKLTAKANRQLQDPLVIMTGNIPTWLT
                     ELGKTCPFFFPFDTRQMLFYVTAFDRDRAMQRLLDTNPEINQSDSQDSRVAPRLDRKKR
                     TVNREELLKQAESVMQDLGSSRAMLEIQYENEVGTGLGPTLEFYALVSQELQRADLGLW
                     RGEEVTLSNPKGSQEGTKYIQNLQGLFALPFGRTAKPAHIAKVKMKFRFLGKLMAKAIM
                     DFRLVDLPLGLPFYKWMLRQETSLTSHDLFDIDPVVARSVYHLEDIVRQKKRLEQDKSQ
                     TKESLQYALETLTMNGCSVEDLGLDFTLPGFPNIELKKGGKDIPVTIHNLEEYLRLVIF
                     WALNEGVSRQFDSFRDGFESVFPLSHLQYFYPEELDQLLCGSKADTWDAKTLMECCRPD
                     HGYTHDSRAVKFLFEILSSFDNEQQRLFLQFVTGSPRLPVGGFRSLNPPLTIVRKTFES
                     TENPDDFLPSVMTCVNYLKLPDYSSIEIMREKLLIAAREGQQSFHLS"
     misc_feature    1817..1822
                     /label=SDS
     misc_feature    3500..3505
                     /label=SDS
     CDS             4065..4130
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      4134..4565
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    4278..4283
                     /label=SDS
     repeat_region   4608..4748
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4608..4737
                     /label=AAV2 ITR
     repeat_region   4609..4749
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4823..5278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5560..5664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5665..6525
                     /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      6696..7284
                     /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 ggtggatggt ggagctgagt
     1501 gccttgtaga aatccgtagc atagtctcag agtcagatgt ttcatcattt gaaatccaac
     1561 atagtggatt tgtgaagcag ctgttgcttt atttgacatc taaaagtgaa aaggatgctg
     1621 tgagcagaga gatcagatta aagcgatttc ttcatgtatt tttttcttct ccacttcctg
     1681 gagaagagcc cattggaaga gtggaaccag tgggtaatgc acctttgttg gcattagttc
     1741 acaagatgaa caactgcctc agccagatgg aacaatttcc agtcaaagta catgatttcc
     1801 ctagtggaaa tgggacagga ggcagctttt ctctcaacag aggatcacag gctttaaaat
     1861 ttttcaacac acatcaatta aaatgccagt tacaaaggca tccagactgt gcaaatgtga
     1921 agcagtggaa gggtggacct gtcaagattg accctctggc tttggtacaa gccatcgaga
     1981 gataccttgt agttagaggg tatggaagag taagagaaga tgatgaagac agcgatgacg
     2041 atggatcaga tgaggaaata gatgagtctc tggctgctca gttcctaaat tcaggaaatg
     2101 taagacacag gctgcagttt tatattggag aacatttgct gccgtataac atgactgtgt
     2161 atcaggcagt acggcagttt agtatacagg ctgaagatga aagagaatcc acagatgatg
     2221 agagcaatcc tctaggcaga gctggtattt ggacaaagac tcatacaata tggtataaac
     2281 ctgtgagaga ggatgaagaa agtaataaag attgtgttgg tggtaaaaga ggaagagccc
     2341 aaacagctcc aacgaaaact tcccctagaa atgcaaaaaa gcatgatgag ttatggcacg
     2401 atggagtgtg cccatcagta tcaaatcctt tagaagttta cctcattccc acaccacctg
     2461 aaaatataac atttgaagac ccgtcattag atgtgatcct tcttttaaga gttttacatg
     2521 ctatcagtcg atactggtat tacttgtatg ataatgcaat gtgcaaggaa attattccaa
     2581 ctagtgaatt tattaacagt aagttaacag caaaagcaaa taggcaactt caagatcctt
     2641 tagtaatcat gacaggaaac atcccaacat ggcttactga gctaggaaaa acctgcccat
     2701 ttttctttcc ttttgatacc cggcaaatgc ttttttatgt aactgcattt gatcgggacc
     2761 gagcaatgca aagattactt gataccaacc cagaaatcaa ccagtctgat tctcaagata
     2821 gcagagttgc acctagattg gatagaaaaa aacgtactgt gaaccgagag gagctgctga
     2881 aacaggcgga gtctgtgatg caggacctcg gcagctcacg ggccatgtta gaaatccagt
     2941 atgaaaatga ggttggtaca ggtcttgggc ctacactgga gttttatgcg cttgtatctc
     3001 aggaactaca gagagctgac ttgggtcttt ggagaggtga agaagtaact cttagcaatc
     3061 caaaagggag ccaagaaggg accaagtata ttcaaaacct ccagggcctg tttgcgcttc
     3121 cctttggtag gacagcaaag ccagctcata tcgcaaaggt taagatgaag tttcgcttct
     3181 taggaaaatt aatggccaag gctatcatgg atttcagatt ggtggacctt ccccttggct
     3241 taccctttta taaatggatg ctacggcaag aaacttcact gacatcacac gatttgtttg
     3301 acatcgaccc agttgtagcc agatcagttt atcacctaga agacattgtc agacagaaga
     3361 aaagacttga acaagataaa tcccagacca aagagagtct acagtatgca ttagaaacct
     3421 tgactatgaa tggctgctca gttgaagatc taggactgga tttcactctg ccagggtttc
     3481 ccaatatcga actgaagaaa ggagggaagg atataccagt cactatccac aatttagagg
     3541 agtatctaag actggttata ttctgggcac taaatgaagg cgtttctagg caatttgatt
     3601 cgttcagaga tggatttgaa tcagtcttcc cactcagtca tcttcagtac ttctacccgg
     3661 aggaactgga tcagctcctt tgtggcagta aagcagacac ttgggatgca aagacactga
     3721 tggaatgctg taggcctgat catggttata ctcatgacag tcgggctgtg aagtttttgt
     3781 ttgagattct cagtagtttt gataatgagc agcagaggtt atttctccag tttgtgactg
     3841 gtagcccaag attgcctgtt ggaggattcc ggagtttgaa tccacctttg acaattgtcc
     3901 gaaagacgtt tgaatcaaca gaaaacccag atgacttctt gccctctgta atgacttgtg
     3961 tgaactatct taagttgccg gactattcaa gcattgagat aatgcgtgaa aaactgttga
     4021 tagcagcaag agaagggcag cagtcgttcc atctttcctg tacagactac aaagaccatg
     4081 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag taactcgaga
     4141 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4201 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4261 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     4321 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4381 cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca tctagcttta
     4441 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4501 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4561 tttaaagatc tggagaccgg tgtggcacgt gcggaccgag cggccgcagg aacccctagt
     4621 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4681 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4741 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4801 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4861 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     5041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     5161 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     5221 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     5281 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5341 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5401 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5461 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5521 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5641 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5701 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5881 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6181 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6241 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6421 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7321 ttttgctggc cttttgctca catgt
//