pAAVLINKv1-pCALM1-Cre-3'-IGF1R-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-IGF1R-FLAG            6780 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 6780)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6780
                     /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..3493
                     /codon_start=1
                     /label=IGF1R
                     /translation="GLAAPAPKLKPRSRPRRRRLNTAKSLRISCTTPSSCPDLKGSGEM
                     SCKWPTPPCPAEAGTPRPQTPTTSPTRKSWRQSTLSLRAEWITRRELSFLTFGLSHCTA
                     SISTAATTRLRSWAAAPPTSSLQGLCPQKEQMTFLGQ*PGSQGLKTPSF*SGRNLRIPM
                     D*F*CMK*NTDHKLRISENVCPDRNTGSMEGPS*TG*TRGTTQPGFRPHLSLGMGRGQI
                     LCSSMSRPKQDMKTSSI*SSLCPSLSC*SWEGW*LCCTSSIEREITAGWGMECCMPL*T
                     RSTSALLMCTFLMSGRWLGRRSP*AGNLGRGRLGWSMKELPRVW*KMNLKPEWPLKQ*T
                     RPQACVRGLSFSTKLL**RSSIVTMWCDCWVWCPKASQHWSSWN**HGAISKVISGL*G
                     QKWRIIQS*HLQA*AR*FRWPERLQTAWHTSTPISSSTETLLPGIAW*PKISQSKSEIL
                     V*REISMRQTITGKEGKGCCPCAGCLLSPSRMESSPLTRTSGPSGSSSGRSPHWPSSPT
                     RACPTSKSFASSWRAAFWTSQTTVLTCCLN*CACAGSITPR*GLPSWRSSAASKRRWSL
                     ASGRSPSTTARRTSCPSRRSWTWSQRTWRASPWTPRPPRPPCHCPTDTQDTRPRTAPAL
                     GCWSSAPASTRDSLTPT*TGAARTSGPCRCPSLRP"
     misc_feature    1530..1535
                     /label=SDS
     misc_feature    1533..1538
                     /label=SDS
     misc_feature    2896..2901
                     /label=SDS
     misc_feature    3318..3323
                     /label=SDS
     CDS             3500..3565
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3569..4000
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3713..3718
                     /label=SDS
     repeat_region   4043..4183
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4043..4172
                     /label=AAV2 ITR
     rep_origin      4258..4713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4995..5099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5100..5960
                     /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      6131..6719
                     /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 gggccttgct gcgcctgccc
     1501 caaaactgaa gccgagaagc aggccgagaa ggaggaggct gaataccgca aagtctttga
     1561 gaatttcctg cacaactcca tcttcgtgcc cagacctgaa aggaagcgga gagatgtcat
     1621 gcaagtggcc aacaccacca tgtccagccg aagcaggaac accacggccg cagacaccta
     1681 caacatcacc gacccggaag agctggagac agagtaccct ttctttgaga gcagagtgga
     1741 taacaaggag agaactgtca tttctaacct tcggcctttc acattgtacc gcatcgatat
     1801 ccacagctgc aaccacgagg ctgagaagct gggctgcagc gcctccaact tcgtctttgc
     1861 aaggactatg cccgcagaag gagcagatga cattcctggg ccagtgacct gggagccaag
     1921 gcctgaaaac tccatctttt taaagtggcc ggaacctgag aatcccaatg gattgattct
     1981 aatgtatgaa ataaaatacg gatcacaagt tgaggatcag cgagaatgtg tgtccagaca
     2041 ggaatacagg aagtatggag gggccaagct aaaccggcta aacccgggga actacacagc
     2101 ccggattcag gccacatctc tctctgggaa tgggtcgtgg acagatcctg tgttcttcta
     2161 tgtccaggcc aaaacaggat atgaaaactt catccatctg atcatcgctc tgcccgtcgc
     2221 tgtcctgttg atcgtgggag ggttggtgat tatgctgtac gtcttccata gaaagagaaa
     2281 taacagcagg ctggggaatg gagtgctgta tgcctctgtg aacccggagt acttcagcgc
     2341 tgctgatgtg tacgttcctg atgagtggga ggtggctcgg gagaagatca ccatgagccg
     2401 ggaacttggg caggggtcgt ttgggatggt ctatgaagga gttgccaagg gtgtggtgaa
     2461 agatgaacct gaaaccagag tggccattaa aacagtgaac gaggccgcaa gcatgcgtga
     2521 gaggattgag tttctcaacg aagcttctgt gatgaaggag ttcaattgtc accatgtggt
     2581 gcgattgctg ggtgtggtgt cccaaggcca gccaacactg gtcatcatgg aactgatgac
     2641 acggggcgat ctcaaaagtt atctccggtc tctgaggcca gaaatggaga ataatccagt
     2701 cctagcacct ccaagcctga gcaagatgat tcagatggcc ggagagattg cagacggcat
     2761 ggcatacctc aacgccaata agttcgtcca cagagacctt gctgcccgga attgcatggt
     2821 agccgaagat ttcacagtca aaatcggaga ttttggtatg acgcgagata tctatgagac
     2881 agactattac cggaaaggag ggaaagggct gctgcccgtg cgctggatgt ctcctgagtc
     2941 cctcaaggat ggagtcttca ccacttactc ggacgtctgg tccttcgggg tcgtcctctg
     3001 ggagatcgcc acactggccg agcagcccta ccagggcttg tccaacgagc aagtccttcg
     3061 cttcgtcatg gagggcggcc ttctggacaa gccagacaac tgtcctgaca tgctgtttga
     3121 actgatgcgc atgtgctggc agtataaccc caagatgagg ccttccttcc tggagatcat
     3181 cagcagcatc aaagaggaga tggagcctgg cttccgggag gtctccttct actacagcga
     3241 ggagaacaag ctgcccgagc cggaggagct ggacctggag ccagagaaca tggagagcgt
     3301 ccccctggac ccctcggcct cctcgtcctc cctgccactg cccgacagac actcaggaca
     3361 caaggccgag aacggccccg gccctggggt gctggtcctc cgcgccagct tcgacgagag
     3421 acagccttac gcccacatga acgggggccg caagaacgag cgggccttgc cgctgcccca
     3481 gtcttcgacc tgctgtacag actacaaaga ccatgacggt gattataaag atcatgacat
     3541 cgactacaag gatgacgatg acaagtaact cgagataatc aacctctgga ttacaaaatt
     3601 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3661 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3721 tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3781 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc
     3901 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3961 tatgtttcag gttcaggggg agatgtggga ggttttttaa agatctggag accggtgtgg
     4021 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4081 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4141 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4201 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4261 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4321 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4381 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4441 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4501 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4561 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4621 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4681 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4741 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4801 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4861 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4921 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4981 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5041 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5101 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5161 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5221 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5281 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5341 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5401 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5461 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5521 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5581 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5641 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5701 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5761 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5821 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5881 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5941 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6001 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6061 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//