pAAVLINKv1-pCALM1-Cre-3'-MED23-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-MED23-FLAG              6663 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 6663)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 19, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6663
                     /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..3376
                     /codon_start=1
                     /label=MED23
                     /translation="AHYRHSSNKIMCLRKAVLI*KKMWRRSIGSGSQ*ATKTTLLPTSL
                     CRAPLLSFFVFSGKCSWKQIILIRLAIEY*RELEPGPW*PM*GHLQISWYMSFLHQQGV
                     SNSINALKFLMTWYGSITLLHWTD*FSAWPCVVTKEMKPRFVIS*FSCCYSNQTILEIE
                     *VTL*RKIPQSTGYRMTGTPST*IITRNIQRSCILRASRNRWILLYRSSLPICPSILGM
                     CVFDSFQYLI**STDF*SCFRYPNHWRLYWII*EAYINFMIVQ*LICITLCTIMKCT*E
                     TAHFSNENSSMRSLAL*RIIDRRAGV*VTLT*NAL*MHERKILGFQMTPTIAD*LAD*S
                     IRWLANLLVPFQTVTGDSMSFPTQLPMLSMLLVWSSWPWQFQAKKLGMPF*MLS*KVSL
                     *CQERTLQHG*MQLV*SSLPYQSHIGLFFMIEL*VSSAAPA*RLKQSGLAIHSASLISL
                     PVISPTLR*VVAIR*LLHMLCGTILASDNFLSFQSFLLKYFFL**RPNSSCFMYTILLD
                     HFYKDFSKRELVV**RLVWRFMTCC*MLTSVAPI*ITWIPSVTSSIT*SICLLVTA*KS
                     K*RRLSVT*NQL*NFVFDSSHTLARWSQLQCLHKP*TVGLQHLSLIRCPCLYQ*L"
     misc_feature    1554..1559
                     /label=SDS
     misc_feature    1628..1633
                     /label=SDS
     misc_feature    2099..2104
                     /label=SDS
     misc_feature    2245..2250
                     /label=SDS
     CDS             3383..3448
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3452..3883
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3596..3601
                     /label=SDS
     repeat_region   3926..4066
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    3926..4055
                     /label=AAV2 ITR
     rep_origin      4141..4596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4983..5843
                     /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      6014..6602
                     /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 ggcccactac aggcattctt
     1501 caaacaaaat aatgtgcctc aggaaagccg ttttaatctg aaaaaaaatg tggaggagga
     1561 gtataggaag tggaagtcaa tgagcaacga aaacgacatt attacccact tctctatgca
     1621 gggctcccct cctctctttc tttgtcttct ctggaaaatg ctcttggaaa cagatcatat
     1681 taatcagatt ggctatagag tattagagag aattggagcc agggccttgg tagcccatgt
     1741 gaggacattt gcagatttcc tggtatatga gttttctaca tcagcagggg gtcagcaact
     1801 caataaatgc attgaaattc ttaatgacat ggtatggaag tataacattg ttacactgga
     1861 cagattaatt ctctgcctgg ccatgcgtag tcacgaagga aatgaagccc aggtttgtta
     1921 tttcataatt cagttgctgt tactcaaacc aaacgatttt agaaatcgag taagtgactt
     1981 tgtgaaggaa aattccccag agcactggtt acagaatgac tggcacacca agcacatgaa
     2041 ttatcacaag aaatatccag agaagttgta ttttgagggc ctcgcggaac aggtggatcc
     2101 tcctgtacag atccagtctc cctatctgcc catctatttt gggaatgtgt gtcttcgatt
     2161 ccttccagta tttgatatag taatccacag atttttagag ttgcttccgg tatccaaatc
     2221 actggagact ctactggatc atctaggagg cttatataaa tttcatgatc gtccagtgac
     2281 ttatctgtat aacactctgc actattatga aatgcacctg agagaccgcg catttctcaa
     2341 acgaaaactc gtccatgcga tcattggctc tctgaaggat aatcgaccgc agggctggtg
     2401 tctaagtgac acttacctga aatgcgctat gaatgcacga gaggaaaatc cttgggttcc
     2461 agatgacacc tactattgca gattgattgg cagactagtc gatacgatgg ctggcaaatc
     2521 tcctggtccc tttccaaact gtgactggag attcaatgag tttcccaacc cagctgccca
     2581 tgctctccat gttacttgtg tggagctcat ggccttggca gtttcaggca aagaagttgg
     2641 gaatgccctt ctaaatgttg tcctaaaaag tcagccttta gtgccaagag agaacattac
     2701 agcatggatg aatgcaattg gtttgatcat cactgcccta ccagagccat attggattgt
     2761 tcttcatgat cgaattgtga gtgtcatcag cagccccagc ttgacgtctg aaacagagtg
     2821 ggttggctat ccattccgcc tctttgattt cactgcctgt catcagtcct actctgagat
     2881 gagttgtagc tatacgttag ctcttgcaca tgctgtgtgg caccattcta gcatcggaca
     2941 actttctctc attccaaagt ttcttactga agtacttctt cctatagtga agaccgaatt
     3001 ccagttgctt tatgtatacc atcttgttgg accattttta caaagatttc agcaagagag
     3061 aactcgttgt atgatagaga ttggtgtggc gttttatgac atgctgctga atgttgacca
     3121 gtgtagcacc catttaaatt acatggatcc catctgtgac ttcctctatc acatgaagta
     3181 tatgtttact ggtgacagcg tgaaagagca agtagagaag attatctgta acttaaaacc
     3241 agctttaaaa cttcgtcttc gattcatcac acacattagc aagatggagc cagctgcagt
     3301 gcctccacaa gccatgaaca gtgggtctcc agcacctcag tctaatcagg tgcccgtgtc
     3361 tttaccagta actcagtgta cagactacaa agaccatgac ggtgattata aagatcatga
     3421 catcgactac aaggatgacg atgacaagta actcgagata atcaacctct ggattacaaa
     3481 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3541 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3601 ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc
     3661 tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt
     3721 tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat
     3781 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3841 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagatctg gagaccggtg
     3901 tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3961 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4021 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4201 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4321 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4441 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4501 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4561 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4621 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4681 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4741 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4801 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgt
//