LOCUS       sequence-379403-        4245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible vector with BsaI cloning site and Cre expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4245)
  AUTHORS   Merle N, Elmshauser S, Strassheimer F, Wanzel M, Konig AM, Funk J, 
            Neumann M, Kochhan K, Helmprobst F, Pagenstecher A, Nist A, 
            Mernberger M, Schneider A, Braun T, Borggrefe T, Savai R, Timofeev 
            O, Stiewe T
  TITLE     Monitoring autochthonous lung tumors induced by somatic CRISPR gene 
            editing in mice using a secreted luciferase.
  JOURNAL   Mol Cancer. 2022 Oct 3;21(1):191. doi: 10.1186/s12943-022-01661-2.
  PUBMED    36192757
REFERENCE   2  (bases 1 to 4245)
  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"; journalName: "Mol 
            Cancer."; date: "2022-10-3"; volume: "21(1):191. doi"; pages: " 
            10.1186/s12943-022-01661-2"
COMMENT     Sequence Label: pShuttle.Cre
FEATURES             Location/Qualifiers
     source          1..4245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        713..1016
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1017..1220
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1171..1191
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1217..1241
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1309..1329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1330..2358
                     /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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     protein_bind    complement(2370..2469)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2486..2505)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2487..2505)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2510..2526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2510..2526)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2628..2647)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2639..3448
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2706..2725)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3595..4183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4084..4103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcacc atggaggcta gagacctaga gcgatcgctc gcggtctcac cgcgttacat
      721 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      781 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      841 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      901 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      961 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1021 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1081 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1141 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1201 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     1261 gctgttttga cctccataga agacaccgac tctagaggat ctaccatgcc caagaagaag
     1321 aggaaggtgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca
     1381 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct
     1441 gagcatacct ggaaaatgct tcagtccgtt tgccggtcgt gggcggcatg gtgcaagttg
     1501 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt
     1561 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt
     1621 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg
     1681 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa
     1741 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata
     1801 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc
     1861 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc
     1921 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact
     1981 aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg
     2041 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact
     2101 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac
     2161 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat
     2221 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta
     2281 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg
     2341 ctggaagatg gcgattaggg aattctagaa cccagctttc ttgtacaaag ttggcattat
     2401 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2461 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2521 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2581 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2641 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2701 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2761 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2821 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2881 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2941 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3001 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3061 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3121 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3181 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3241 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3301 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3361 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3421 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3481 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3541 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3601 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3661 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3721 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3781 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3841 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3901 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3961 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4021 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4081 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4141 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4201 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
