LOCUS       sequence-349298-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre-recombinase and can be packaged into AAV particles..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  AUTHORS   Kaltenbacher T, Loprich J, Maresch R, Weber J, Muller S, Oellinger 
            R, Gross N, Griger J, de Andrade Kratzig N, Avramopoulos P, 
            Ramanujam D, Brummer S, Widholz SA, Barthel S, Falcomata C, Pfaus A,
            Alnatsha A, Mayerle J, Schmidt-Supprian M, Reichert M, Schneider G, 
            Ehmer U, Braun CJ, Saur D, Engelhardt S, Rad R
  TITLE     CRISPR somatic genome engineering and cancer modeling in the mouse 
            pancreas and liver.
  JOURNAL   Nat Protoc. 2022 Apr;17(4):1142-1188. doi: 
            10.1038/s41596-021-00677-0. Epub 2022 Mar 14.
  PUBMED    35288718
REFERENCE   2  (bases 1 to 6250)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41596-021-00677-0"; journalName: "Nat Protoc"; date: 
            "2022-04"; volume: "17"; issue: "4"; pages: "1142-1188"
COMMENT     Sequence Label: scAAV-Cre
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        196..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     700..724
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          839..971
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     954..973
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1016..1034
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1073..1093
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2140..2274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2164..2183)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2218..2237
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2424..2553
                     /label=AAV2 ITR
     rep_origin      2924..3437
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3221..3242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3919..3938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4038..4060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4098..4116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4184..4288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4289..4357,4358..5149)
                     /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"
     primer_bind     complement(4507..4526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5320..5908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5809..5828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaaagcttc agtctcctga gtagctggga ctacaggagc ttgttaccac acccagctcc
       61 agtttataaa ttcatctcca gtttataaag gaggaaaccg aggtactgag aggttaaaaa
      121 accttcctgc agacacgcgt cttaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
      721 gatcactaga agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc
      781 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
      841 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
      901 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
      961 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     1021 cgactcacta taggctagcc tcgacggtac cgcgggcccg ggatccatgg tgcccaagaa
     1081 gaagaggaaa gtctccaacc tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga
     1141 tgccacctct gatgaagtca ggaagaacct gatggacatg ttcagggaca ggcaggcctt
     1201 ctctgaacac acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa
     1261 gctgaacaac aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta
     1321 cctgcaagcc agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat
     1381 gctgcacagg agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat
     1441 gaggagaatc agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt
     1501 tgaacgcact gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga
     1561 catcaggaac ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat
     1621 tgccagaatc agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat
     1681 tggcaggacc aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt
     1741 taccaagctg gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta
     1801 cctgttctgc cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc
     1861 cacccgggcc ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga
     1921 tgactctggg cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccag
     1981 ggacatggcc agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa
     2041 tgtgaacatt gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag
     2101 gctgctcgag gatggggact gagcggccgc caccgcgggg atccagacat gataagatac
     2161 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2221 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2281 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttcggat
     2341 cctctagagt cgacctgcag gcatgcaagc tctagagcat ggctacgtag ataagtagca
     2401 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     2461 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     2521 ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag aggcccgcac
     2581 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc gttgcaatgg
     2641 ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt tcttctactc
     2701 aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat ttgcgtgatg
     2761 gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag gattctggcg
     2821 taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc tctgattcta
     2881 acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc ctgtagcggc
     2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3361 atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt ggggcttttc
     3421 tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc gttcatcgat
     3481 tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga gacctctcaa
     3541 aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat catattgatg
     3601 gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca cattactcag
     3661 gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt gaaataaagg
     3721 cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat ttagctttat
     3781 gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat gatttattgg
     3841 atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3901 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3961 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4021 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4081 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4141 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagttcctt tttttctgcg
     5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5941 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6001 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6061 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6121 gcgttggccg attcattaat gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     6181 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     6241 gagggagtgg
//
