pAAVLINKv1-pCALM1-Cre-3'-POLR3A-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-POLR3A-FLAG            6579 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 6579)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6579
                     /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..3292
                     /codon_start=1
                     /label=POLR3A
                     /translation="VPSLTYHR*LPVWDSRPSVALECQTALKTGPCLILKNTQSSQLPK
                     ALWLIAFIPV*HQLSFSSTQWPAGKV*STRL*RQLKRDTCSEGLSNLLKIFAPSMI*QS
                     EALLAILSSSFMEEMA*ILQLWREKMNLWSLKGFWTTSKQSSRVPVSLLSAKTS*S*PQ
                     SPS*RRVSSSAARTASCRK*KNSLRGSLRRSRKPEINMASMITAQQSPVCCTSWTASPP
                     PK*KSFWRPVGTST*GHRWSQVLQWVLCVPRALVSQAPR*P*RLSTLQVWPP*TSPWAC
                     PGLKRSSTLPRPSALQLSQHS*TRMTTRIMLAS*KGELRKPSWERFPSILKKCFFLMTA
                     LFSSSSPWNGLGF*DWK*TLRQ*DIPSAHPSSV*SPVMWLFMVRLWCVSPPERTARAPC
                     TTCCSS*KRISPRWWCRAFQRCPELSSTLTSRVERRSTSFWWKVITCGQSWPHTV*RAP
                     EPPPITPMRWRKLWASRPPGQRSSMKSSTPW*TTA*ASTGGT*CCSPTS*PTRVKSWAS
                     LGLAWPR*RRVC*CWPPLRRRLTISLTLPTSGRRTLCVGCLSASSWESQ*TLEPGSSSC
                     FTRLTGTRTLPRGP*SSTQMNSTSPLS"
     misc_feature    2972..2977
                     /label=SDS
     misc_feature    3078..3083
                     /label=SDS
     CDS             3299..3364
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3368..3799
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3512..3517
                     /label=SDS
     repeat_region   3842..3982
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    3842..3971
                     /label=AAV2 ITR
     rep_origin      4057..4512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4794..4898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4899..5759
                     /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      5930..6518
                     /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 ggttccttca ttaacatatc
     1501 acagatgatt gcctgtgtgg gacagcaggc catcagtggc tctcgagtgc cagacggctt
     1561 tgaaaacagg tccttgcctc attttgaaaa acactcaaag ctcccagctg ccaaaggctt
     1621 tgtggctaat agcttttatt ccggtttgac accaactgag tttttcttcc acacaatggc
     1681 cggccgggaa ggtctagtcg acacggctgt aaagacagct gaaacgggat acatgcagcg
     1741 aaggcttgtc aaatctcttg aagatctttg ctcccagtat gatctgacag tccgaagctc
     1801 tactggcgat attatccagt tcatttatgg aggagatggc ttagatcctg cagctatgga
     1861 gggaaaagat gaacctttgg agtttaaaag ggttctggac aacatcaaag cagtcttccc
     1921 gtgtcccagt gagcctgctc tcagcaaaaa cgagctgatc ctgaccacag agtccatcat
     1981 gaagaagagt gagttcctct gctgccagga cagcttcctg caggaaataa aaaaattcat
     2041 taagggggtc tctgagaaga tcaagaaaac cagagataaa tatggcatca atgataacgg
     2101 cacaacagag ccccgtgtgc tgtaccagct ggaccgcatc acccccaccc aagtagaaaa
     2161 gtttctggag acctgtaggg acaagtacat gagggcacag atggagccag gttctgcagt
     2221 gggtgctctg tgtgcccaga gcattggtga gccaggcacc cagatgaccc tgaagacttt
     2281 ccactttgca ggtgtggcct ccatgaacat caccctgggc gtgccccgga ttaaagagat
     2341 catcaacgct tccaaggcca tcagcactcc aattatcaca gcacagctag acaaggatga
     2401 cgacgcggat tatgctcgcc tcgtgaaagg gagaattgag aaaaccctct tgggagagat
     2461 ttccgagtat attgaagaag tgtttcttcc tgatgactgc tttattctcg tcaagctctc
     2521 cctggaacgg attaggcttc tgagactgga agtgaacgct gagacagtga gatattccat
     2581 ctgcacatcc aagctccgtg tgaagcccgg tgatgtggct gttcatggtg aggctgtggt
     2641 gtgtgtcacc cccagagaga acagcaagag ctccatgtac tacgtgctgc agttcctgaa
     2701 agaggatctc cccaaggtgg tggtgcaggg cattccagag gtgtccagag ctgtcatcca
     2761 cattgacgag cagagtggaa aggagaagta caagcttctg gtggaaggtg ataacctgcg
     2821 ggcagtcatg gccacacacg gtgtgaaggg cacccgaacc acctccaata acacctatga
     2881 ggtggagaaa actctgggca tcgaggccgc ccggacaacg atcatcaatg aaatccagta
     2941 caccatggtg aaccacggca tgagcatcga caggaggcac gtgatgctgc tctccgacct
     3001 catgacctac aagggtgaag tcctgggcat cactaggttt ggcctggcca agatgaagga
     3061 gagtgtgctg atgctggcct cctttgagaa gacggctgac catctctttg acgctgccta
     3121 cttcgggcag aaggactctg tgtgtggggt gtctgagtgc atcatcatgg gaatcccaat
     3181 gaacattgga accgggctct tcaagctgct tcacaaggct gacagggacc cgaaccctcc
     3241 caagaggccc ctgatcttcg acacaaatga attccacatc ccccttgtca catgtacaga
     3301 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     3361 caagtaactc gagataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3421 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3481 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     3541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3601 ggcactgaca attccgtggt gtttatttgt gaaatttgtg atgctattgc tttatttgta
     3661 accatctagc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3721 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3781 gatgtgggag gttttttaaa gatctggaga ccggtgtggc acgtgcggac cgagcggccg
     3841 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3901 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3961 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4021 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4081 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4141 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4201 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4261 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4321 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4381 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4441 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4501 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4561 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4621 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4681 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4921 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4981 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5041 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5101 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5161 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5221 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5281 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5341 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5401 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5461 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5521 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5581 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5641 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5701 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5761 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6541 tacggttcct ggccttttgc tggccttttg ctcacatgt
//