LOCUS       sequence-345275-        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        302..605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     760..780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     806..830
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          945..1077
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1060..1079
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1122..1140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1179..1199
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2246..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2324..2343
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2530..2659
                     /label=AAV2 ITR
     rep_origin      3030..3543
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3117..3136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3327..3348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4025..4044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4144..4166
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4204..4222)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4290..4394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4395..4463,4464..5255)
                     /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(4613..4632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5426..6014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5915..5934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggtaaa agcttcagtc
      121 tcctgagtag ctgggactac aggagcttgt taccacaccc agctccagtt tataaattca
      181 tctccagttt ataaaggagg aaaccgaggt actgagaggt taaaaaacct tcctgcagac
      241 acgcgtctta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      481 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      541 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      601 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
      661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      721 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
      781 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc actagaagct
      841 ttattgcggt agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag
      901 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
      961 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     1021 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     1081 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
     1141 ctagcctcga cggtaccgcg ggcccgggat ccatggtgcc caagaagaag aggaaagtct
     1201 ccaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc acctctgatg
     1261 aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct gaacacacct
     1321 ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg aacaacagga
     1381 aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg caagccagag
     1441 gcctggctgt gaagaccatc caacagcacc tgggccagct caacatgctg cacaggagat
     1501 ctggcctgcc tcgcccttct gactccaatg ctgtgtccct ggtgatgagg agaatcagaa
     1561 aggagaatgt ggatgctggg gagagagcca agcaggccct ggcctttgaa cgcactgact
     1621 ttgaccaagt cagatccctg atggagaact ctgacagatg ccaggacatc aggaacctgg
     1681 ccttcctggg cattgcctac aacaccctgc tgcgcattgc cgaaattgcc agaatcagag
     1741 tgaaggacat ctcccgcacc gatggtggga gaatgctgat ccacattggc aggaccaaga
     1801 ccctggtgtc cacagctggt gtggagaagg ccctgtccct gggggttacc aagctggtgg
     1861 agagatggat ctctgtgtct ggtgtggctg atgaccccaa caactacctg ttctgccggg
     1921 tcagaaagaa tggtgtggct gccccttctg ccacctccca actgtccacc cgggccctgg
     1981 aagggatctt tgaggccacc caccgcctga tctatggtgc caaggatgac tctgggcaga
     2041 gatacctggc ctggtctggc cactctgcca gagtgggtgc tgccagggac atggccaggg
     2101 ctggtgtgtc catccctgaa atcatgcagg ctggtggctg gaccaatgtg aacattgtga
     2161 tgaactacat cagaaacctg gactctgaga ctggggccat ggtgaggctg ctcgaggatg
     2221 gggactgagc ggccgccacc gcggggatcc agacatgata agatacattg atgagtttgg
     2281 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2341 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2401 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgac
     2461 ctgcaggcat gcaagctcta gagcatggct acgtagataa gtagcatggc gggttaatca
     2521 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2581 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2641 tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2701 aacagttgcg cagcctgaat ggcgaatggc gattccgttg caatggctgg cggtaatatt
     2761 gttctggata ttaccagcaa ggccgatagt ttgagttctt ctactcaggc aagtgatgtt
     2821 attactaatc aaagaagtat tgcgacaacg gttaatttgc gtgatggaca gactctttta
     2881 ctcggtggcc tcactgatta taaaaacact tctcaggatt ctggcgtacc gttcctgtct
     2941 aaaatccctt taatcggcct cctgtttagc tcccgctctg attctaacga ggaaagcacg
     3001 ttatacgtgc tcgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3061 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3481 aatttaaata tttgcttata caatcttcct gtttttgggg cttttctgat tatcaaccgg
     3541 ggtacatatg attgacatgc tagttttacg attaccgttc atcgattctc ttgtttgctc
     3601 cagactctca ggcaatgacc tgatagcctt tgtagagacc tctcaaaaat agctaccctc
     3661 tccggcatga atttatcagc tagaacggtt gaatatcata ttgatggtga tttgactgtc
     3721 tccggccttt ctcacccgtt tgaatcttta cctacacatt actcaggcat tgcatttaaa
     3781 atatatgagg gttctaaaaa tttttatcct tgcgttgaaa taaaggcttc tcccgcaaaa
     3841 gtattacagg gtcataatgt ttttggtaca accgatttag ctttatgctc tgaggcttta
     3901 ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt tattggatgt tggaatcgcc
     3961 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     4021 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4081 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4141 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4201 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4261 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4321 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4381 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4441 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4501 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4561 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4621 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4681 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4741 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4801 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4861 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4921 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4981 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5041 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5101 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5161 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5221 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5281 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5341 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5401 ccgtagaaaa gatcaaagga tcttcttgag ttcctttttt tctgcgcgta atctgctgct
     5461 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5521 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5581 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5641 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5701 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5761 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5821 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5881 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5941 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6001 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6061 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6121 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6181 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6241 attaatgcag
//
