pAAVLINKv1-pCALM1-Cre-3'-TNRC6B-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-TNRC6B-FLAG             7186 bp DNA     circular SYN 17-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7186)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7186
                     /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..3899
                     /codon_start=1
                     /label=TNRC6B
                     /translation="GSASSHNSASWGQGGKKQMKCSLKGGNNDSWMNPLAKQFSNMGLL
                     SQTEDNPSSKMDLSVGSLSDKKFDVDKRAMNLGDFNDIMRKDRSGFRPPNSKDMGTTDS
                     GPYFEKLTLPFSNQDGCLGDEAPCSPFSPSPSYKLSPSGSTLPNVSLGAIGTGLNPQNF
                     AARQGGSHGLFGNSTAQSRGLHTPVQPLNSSPSLRAQVPPQFISPQVSASMLKQFPNSG
                     LSPGLFNVGPQLSPQQIAMLSQLPQIPQFQLACQLLLQQQQQQQLLQNQRKISQAVRQQ
                     QEQQLARMVSALQQQQQQQQRQPGMKHSPSHPVGPKPHLDNMVPNALNVGLPDLQTKGP
                     IPGYGSGFSSGGMDYGMVGGKEAGTESRFKQWTSMMEGLPSVATQEANMHKNGAIVAPG
                     KTRGGSPYNQFDIIPGDTLGGHTGPAGDSWLPAKSPPTNKIGSKSSNASWPPEFQPGVP
                     WKGIQNIDPESDPYVTPGSVLGGTATSPIVDTDHQLLRDNTTGSNSSLNTSLPSPGAWP
                     YSASDNSFTNVHSTSAKFPDYKSTWSPDPIGHNPTHLSNKMWKNHISSRNTTPLPRPPP
                     GLTNPKPSSPWSSTAPRSVRGWGTQDSRLASASTWSDGGSVRPSYWLVLHNLTPQIDGS
                     TLRTICMQHGPLLTFHLNLTQGTALIRYSTKQEAAKAQTALHMCVLGNTTILAEFATDD
                     EVSRFLAQAQPPTPAATPSAPAAGWQSLETGQNQSDPVGPALNLFGGSTGLGQWSSSAG
                     GSSGADLAGASLWGPPNYSSSLWGVPTVEDPHRMGSPAPLLPGDLLGGGSDSI"
     misc_feature    1520..1525
                     /label=SDS
     misc_feature    1553..1558
                     /label=SDS
     misc_feature    2563..2568
                     /label=SDS
     misc_feature    3204..3209
                     /label=SDS
     misc_feature    3481..3486
                     /label=SDS
     misc_feature    3881..3886
                     /label=SDS
     CDS             3906..3971
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3975..4406
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    4119..4124
                     /label=SDS
     repeat_region   4449..4589
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4449..4578
                     /label=AAV2 ITR
     rep_origin      4664..5119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5401..5505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5506..6366
                     /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      6537..7125
                     /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 gggcagtgct tcctcccaca
     1501 actcagcaag ctggggacaa ggaggaaaga aacaaatgaa gtgctcactc aaaggaggaa
     1561 acaatgattc atggatgaat cctcttgcca aacagttttc aaatatggga ttgctgagtc
     1621 agactgaaga taatccaagc agcaaaatgg atttgtctgt aggaagcctt tcagataaaa
     1681 aatttgatgt ggacaagcga gcgatgaatc tcggggattt taatgatatc atgaggaagg
     1741 atcgatctgg gttccgtcca cctaattcca aagacatggg aaccacagat agtgggcctt
     1801 attttgagaa gctgactttg cctttctcca atcaagatgg gtgccttggg gatgaggctc
     1861 cctgctctcc cttctcccct tctcccagct acaagctgtc tccctctggt tccacactac
     1921 ccaacgtcag ccttggagca atcggcacag ggctcaaccc ccaaaacttc gctgctagac
     1981 aaggcggtag tcatggtttg tttggaaaca gcacagcaca atcgagaggt ctgcacacac
     2041 ccgtgcagcc actaaattct tctcccagtc tccgggcgca agtgcctccc cagtttattt
     2101 ccccccaggt ttctgcctca atgctcaagc agtttcccaa cagtggcctg agtccaggtc
     2161 ttttcaatgt ggggccccag ttatctcctc aacaaattgc catgctgagc cagcttccac
     2221 aaattcccca gtttcagttg gcatgtcagc ttctcttgca gcagcagcaa cagcagcagt
     2281 tgttacagaa ccagagaaag atttctcaag ctgtacgcca acagcaagag cagcagctgg
     2341 ctcgaatggt gagtgcactg cagcagcagc agcagcagca gcagaggcag ccaggcatga
     2401 agcactcgcc ctctcatcct gttgggccca agccgcatct ggacaacatg gtacccaacg
     2461 cattgaatgt ggggctccca gaccttcaaa ccaaagggcc aatacctgga tatggttctg
     2521 gcttcagctc tggcggcatg gactatggca tggttggtgg gaaggaggct ggaaccgagt
     2581 ctcgctttaa acagtggacc tccatgatgg aggggctgcc ctctgtagcc acacaggaag
     2641 ccaatatgca caaaaatggc gctatagtgg cccctggtaa aacccgggga gggtcaccgt
     2701 acaaccagtt tgatatcatc cctggtgaca cactgggtgg ccatacgggt cctgctggtg
     2761 atagctggtt acctgccaaa tctccaccaa caaataaaat cggaagtaaa tccagcaatg
     2821 ccagttggcc tccagaattc caaccaggag tgccatggaa aggtatccaa aacattgacc
     2881 ctgaatctga cccctatgtc accccaggaa gtgtgctggg gggtacagcc acatctccca
     2941 ttgtagatac tgaccaccaa ctgctgcggg ataacaccac agggtctaat tcttccctca
     3001 acacctcgct gccttcacct ggtgcctggc cctacagtgc ctctgacaac tcctttacca
     3061 acgttcatag cacttcagca aagttccctg attacaaatc aacatggtcc ccagatccca
     3121 taggacacaa ccccactcat ctctccaaca agatgtggaa aaaccatatt tcctccagga
     3181 acactacacc gctgccccgc ccacctcctg gtctgaccaa ccccaaacca tcatctccct
     3241 ggagcagcac agcaccccga tcagtcaggg ggtgggggac acaggactca cggctcgcct
     3301 cggcctctac ctggagtgat ggtggctcag ttcgtcctag ttactggctg gttcttcaca
     3361 atctcacccc acagattgat gggtcaacct tgagaacgat ctgcatgcag catggcccac
     3421 tgctgacatt ccatctgaat ctaacccagg gcactgccct gatccgatac agcaccaaac
     3481 aggaggcggc caaggcccaa actgcactgc acatgtgtgt gttgggaaac actaccatcc
     3541 ttgctgagtt tgccactgat gatgaagtca gccgctttct ggcacaagct cagcccccta
     3601 cacctgcagc aaccccaagt gcgccagctg cggggtggca gtcgctggag accggccaga
     3661 accagtcaga tcccgtggga cctgctctga atctttttgg tgggtccact gggctcgggc
     3721 agtggagcag cagcgctggt ggcagcagcg gggccgatct tgctggcgct tcattgtggg
     3781 ggcccccaaa ctattcttct agcttatggg gagtcccaac ggtggaagat ccccatagga
     3841 tgggcagccc tgctccttta ctacctggtg accttctggg aggagggtcg gattcaatct
     3901 gtacagacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3961 acgatgacaa gtaactcgag ataatcaacc tctggattac aaaatttgtg aaagattgac
     4021 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4081 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4141 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4201 gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt
     4261 atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4321 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4381 agggggagat gtgggaggtt ttttaaagat ctggagaccg gtgtggcacg tgcggaccga
     4441 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4501 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4561 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4621 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4741 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4861 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4981 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5041 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5101 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5161 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5221 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5281 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5341 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//