pAAVLINKv1-pCALM1-Cre-3'-OTOF-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-OTOF-FLAG               7264 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 7264)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7264
                     /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..3977
                     /codon_start=1
                     /gene="OTOF"
                     /gene_synonym="AUNB1; DFNB6; DFNB9; FER1L2; NSRD9"
                     /product="otoferlin isoform a"
                     /label=OTOF
                     /note="isoform a is encoded by transcript variant 
                     1;fer-1-like protein 2; fer-1-like family member 2"
                     /db_xref="GI:34740331"
                     /db_xref="CCDS:CCDS1725.1"
                     /db_xref="GeneID:9381"
                     /db_xref="HGNC:HGNC:8515"
                     /db_xref="MIM:603681"
                     /protein_id="NP_919224.1"
                     /translation="GVQSSLIHNYKKNPNFNTLVKWFEVDLPENELLHPPLNIRVVDCR
                     AFGRYTLVGSHAVSSLRRFIYRPPDRSAPSWNTTVRLLRRCRVLCNGGSSSHSTGEVVV
                     TMEPEVPIKKLETMVKLDATSEAVVKVDVAEEEKEKKKKKKGTAEEPEEEEPDESMLDW
                     WSKYFASIDTMKEQLRQQEPSGIDLEEKEEVDNTEGLKGSMKGKEKARAAKEEKKKKTQ
                     SSGSGQGSEAPEKKKPKIDELKVYPKELESEFDNFEDWLHTFNLLRGKTGDDEDGSTEE
                     ERIVGRFKGSLCVYKVPLPEDVSREAGYDSTYGMFQGIPSNDPINVLVRVYVVRATDLH
                     PADINGKADPYIAIRLGKTDIRDKENYISKQLNPVFGKSFDIEASFPMESMLTVAVYDW
                     DLVGTDDLIGETKIDLENRFYSKHRATCGIAQTYSTHGYNIWRDPMKPSQILTRLCKDG
                     KVDGPHFGPPGRVKVANRVFTGPSEIEDENGQRKPTDEHVALLALRHWEDIPRAGCRLV
                     PEHVETRPLLNPDKPGIEQGRLELWVDMFPMDMPAPGTPLDISPRKPKKYELRVIIWNT
                     DEVVLEDDDFFTGEKSSDIFVRGWLKGQQEDKQDTDVHYHSLTGEGNFNWRYLFPFDYL
                     AAEEKIVISKKESMFSWDETEYKIPARLTLQIWDADHFSADDFLGAIELDLNRFPRGAK
                     TAKQCTMEMATGEVDVPLVSIFKQKRVKGWWPLLARNENDEFELTGKVEAELHLLTAEE
                     AEKNPVGLARNEPDPLEKPNRPDTSFIWFLNPLKSARYFLWHTYRWLLLKLLLLLLLLL
                     LLALFLYSVPGYLVKKILGA"
     misc_feature    1762..1767
                     /label=SDS
     misc_feature    1885..1890
                     /label=SDS
     misc_feature    1936..1941
                     /label=SDS
     misc_feature    1939..1944
                     /label=SDS
     misc_feature    2053..2058
                     /label=SDS
     misc_feature    2320..2325
                     /label=SDS
     misc_feature    2632..2637
                     /label=SDS
     misc_feature    3295..3300
                     /label=SDS
     misc_feature    3659..3664
                     /label=SDS
     misc_feature    3736..3741
                     /label=SDS
     misc_feature    3914..3919
                     /label=SDS
     misc_feature    3917..3922
                     /label=SDS
     CDS             3984..4049
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      4053..4484
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    4197..4202
                     /label=SDS
     repeat_region   4527..4667
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4527..4656
                     /label=AAV2 ITR
     repeat_region   4528..4668
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4742..5197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5479..5583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5584..6444
                     /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      6615..7203
                     /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 gggggtgcag tcgtccctga
     1501 tccacaatta taagaagaac cccaacttca acaccctcgt caagtggttt gaagtggacc
     1561 tcccagagaa cgagctgctg cacccgccct tgaacatccg tgtggtggac tgccgggcct
     1621 tcggtcgcta cacactggtg ggctcccatg ccgtcagctc cctgcgacgc ttcatctacc
     1681 ggcccccaga ccgctcggcc cccagctgga acaccacggt caggcttctc cggcgctgcc
     1741 gtgtgctgtg caatgggggc tcctcctctc actccacagg ggaggttgtg gtgactatgg
     1801 agccagaggt acccatcaag aaactggaga ccatggtgaa gctggacgcg acttctgaag
     1861 ctgttgtcaa ggtggatgtg gctgaggagg agaaggagaa gaagaagaag aagaagggca
     1921 ctgcggagga gccagaggag gaggagccag acgagagcat gctggactgg tggtccaagt
     1981 actttgcctc cattgacacc atgaaggagc aacttcgaca acaagagccc tctggaattg
     2041 acttggagga gaaggaggaa gtggacaata ccgagggcct gaaggggtca atgaagggca
     2101 aggagaaggc aagggctgcc aaagaggaga agaagaagaa aactcagagc tctggctctg
     2161 gccaggggtc cgaggccccc gagaagaaga aacccaagat tgatgagctt aaggtatacc
     2221 ccaaagagct ggagtccgag tttgataact ttgaggactg gctgcacact ttcaacttgc
     2281 ttcggggcaa gaccggggat gatgaggatg gctccaccga ggaggagcgc attgtgggac
     2341 gcttcaaggg ctccctctgc gtgtacaaag tgccactccc agaggacgtg tcccgggaag
     2401 ccggctacga ctccacctac ggcatgttcc agggcatccc gagcaatgac cccatcaatg
     2461 tgctggtccg agtctatgtg gtccgggcca cggacctgca ccctgctgac atcaacggca
     2521 aagctgaccc ctacatcgcc atccggctag gcaagactga catccgcgac aaggagaact
     2581 acatctccaa gcagctcaac cctgtctttg ggaagtcctt tgacatcgag gcctccttcc
     2641 ccatggaatc catgctgacg gtggctgtgt atgactggga cctggtgggc actgatgacc
     2701 tcattgggga aaccaagatc gacctggaga accgcttcta cagcaagcac cgcgccacct
     2761 gcggcatcgc ccagacctac tccacacatg gctacaatat ctggcgggac cccatgaagc
     2821 ccagccagat cctgacccgc ctctgcaaag acggcaaagt ggacggcccc cactttgggc
     2881 cccctgggag agtgaaggtg gccaaccgcg tcttcactgg gccctctgag attgaggacg
     2941 agaacggtca gaggaagccc acagacgagc atgtggcgct gttggccctg aggcactggg
     3001 aggacatccc ccgcgcaggc tgccgcctgg tgccagagca tgtggagacg aggccgctgc
     3061 tcaaccccga caagccgggc atcgagcagg gccgcctgga gctgtgggtg gacatgttcc
     3121 ccatggacat gccagcccct gggacgcctc tggacatctc acctcggaag cccaagaagt
     3181 acgagctgcg ggtcatcatc tggaacacag atgaggtggt cttggaggac gacgacttct
     3241 tcacagggga gaagtccagt gacatcttcg tgagggggtg gctgaagggc cagcaggagg
     3301 acaagcagga cacagacgtc cactaccact ccctcactgg cgagggcaac ttcaactggc
     3361 gctacctgtt ccccttcgac tacctggcgg cggaggagaa gatcgtcatc tccaagaagg
     3421 agtccatgtt ctcctgggac gagaccgagt acaagatccc cgcgcggctc accctgcaga
     3481 tctgggatgc ggaccacttc tccgctgacg acttcctggg ggccatcgag ctggacctga
     3541 accggttccc gcggggcgca aagacagcca agcagtgcac catggagatg gccaccgggg
     3601 aggtggacgt gcccctcgtg tccatcttca agcaaaagcg cgtcaaaggc tggtggcccc
     3661 tcctggcccg caatgagaac gatgagtttg agctcacggg caaggtggag gctgagctgc
     3721 atttactgac agcagaggag gcagagaaga acccagtggg cctggcccgc aatgaacctg
     3781 accccctaga gaaacccaac cggcccgaca cgagcttcat ctggttcctg aaccctctca
     3841 agtcggctcg ctacttcttg tggcacacgt atcgctggct gctcctcaaa ctgttgctgc
     3901 tcctgctgct gctcctcctc ctcgccctgt tcctctactc tgtgcctggc tacctggtca
     3961 agaaaatcct cggggcctgt acagactaca aagaccatga cggtgattat aaagatcatg
     4021 acatcgacta caaggatgac gatgacaagt aactcgagat aatcaacctc tggattacaa
     4081 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4141 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4201 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     4261 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta tttgtgaaat
     4321 ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt tgtgatgcta
     4381 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4441 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagatct ggagaccggt
     4501 gtggcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4561 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4621 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4681 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4801 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4921 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5041 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     5101 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     5161 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     5221 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5281 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5341 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5401 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5461 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5521 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5581 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5641 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5701 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5761 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5821 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5881 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5941 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6001 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6061 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6121 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6181 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6241 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6301 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6361 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6421 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6601 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6781 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7201 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7261 atgt
//