LOCUS       Exported                5171 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Donor vector for mFTH1 knock-in via landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    pBGO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5171)
  AUTHORS   Ohtsuka M, Miura H, Mochida K, Hirose M, Hasegawa A, Ogura A, 
            Mizutani R, Kimura M, Isotani A, Ikawa M, Sato M, Gurumurthy CB
  TITLE     One-step generation of multiple transgenic mouse lines using an 
            improved Pronuclear Injection-based Targeted Transgenesis (i-PITT).
  JOURNAL   BMC Genomics. 2015 Apr 9;16:274. doi: 10.1186/s12864-015-1432-5.
  PUBMED    25887549
REFERENCE   2  (bases 1 to 5171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5171
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    161..194
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    203..236
                     /label=loxJTZ17
                     /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)."
     primer_bind     620..639
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    638..693
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     739..758
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1397..1423)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1469..1488)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1496..2068
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    complement(3307..3354)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    3355..3388
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     primer_bind     complement(3472..3488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      3536..3567
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3568..3670
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3671..4486
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3744..3763)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4510..4537
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4549..5136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4629..4648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcgg cctcggatcc gcgatcgatc cgcggtgcgg gtgccagggc gtgcccttgg
      181 gctccccggg cgcgtactcc acataacttc gtatagcata cattatagca atttatcgaa
      241 gttcctattc cgaagttcct attctatcag aagtatagga acttcaccgg taagcttggg
      301 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
      361 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
      421 aaataacaca aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa
      481 taaggaatgg acagcagggg gctgtttcat atactgatga cctctttata gccacctttg
      541 ttcatggcag ccagcatatg gcatatgttg ccaaactcta aaccaaatac tcattctgat
      601 gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac
      661 acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct
      721 tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca
      781 gggcattggc cacaccagcc accaccttct gataggcagc ctgcacctga ggagtgaatt
      841 cctcgagtta gctctcatca ccgtgtccca gggtgtgctt gtcaaagaga tattctgcca
      901 tgccagcttc aggggcaccc atcttgcgta agttggtcac gtggtcaccc agttctttaa
      961 tggatttcac ctgttcactc agataatacg tctcaatgaa gtcacataag tggggatcat
     1021 tcttgtcagt agccagtttg tgcagttcca gtagtgactg attcacactc ttttccaagt
     1081 gcagtgcaca ctccattgca ttcagcccgc tctcccagtc atcacggtct ggtttcttta
     1141 tatcctgcag gaagattcgg ccacctcgct ggttctgcag cttcatcagt ttctcggcat
     1201 gctccctctc ctcatgagat tggtggagaa agtatttggc aaagttcttc agagccacat
     1261 catctcggtc aaaataacaa gacatagaca gatagacgta ggaggcatac aactccaggt
     1321 tgatctggcg gttgatggca gcctccgcgt cctggtggta gttctggcgc acttgcgagg
     1381 gagacgcggt ggtcatagcg taatctggaa catcgtatgg gtacatggtg gcactcgagg
     1441 tcaattcttt gccaaaatga tgagacagca caataaccag cacgttgccc aggagctgta
     1501 ggaaaaagaa gaaggcatga acatggttag cagaggggcc cggtttggac tcagagtatt
     1561 ttatcctcat ctcaaacagt gtatatcatt gtaaccataa agagaaaggc aggatgatga
     1621 ccaggatgta gttgtttcta ccaataagaa tatttccacg ccagccagaa tttatatgca
     1681 gaaatattct accttatcat ttaattataa caattgttct ctaaaactgt gctgaagtac
     1741 aatataatat accctgattg ccttgaaaaa aaagtgatta gagaaagtac ttacaatctg
     1801 acaaataaac aaaagtgaat ttaaaaattc gttacaaatg caagctaaag tttaacgaaa
     1861 aagttacaga aaatgaaaag aaaataagag gagacaatgg ttgtcaacag agtagaaagt
     1921 gaaagaaaca aaattatcat gagggtccat ggtgatacaa gggacatctt cccattctaa
     1981 acaacaccct gaaaactttg ccccctccat ataacatgaa ttttacaata gcgaaaaaga
     2041 aagaacaatc aagggtcccc aaactcaccc tgaagttctc aggatcgggg gagcgggcgg
     2101 ggcagtgagt caggaggggt agcagggcgg ggccgggctg ggtggggcgg ggcttgcgtc
     2161 gtgctcacca atggccgcgc cacttctccc tggggaaatg agtcaccgct ccagactggt
     2221 ttaagggatt gtacggttta aaccgccccc aggtgtcatc actccctgac ggcgctcacc
     2281 gactataaaa atcatcttta atggaagact ggtccagaac ccattgctcc cgttacataa
     2341 tggggaaggt gagaaaatga aaggttgcct aatggagcga gaaagcaatt gctgaaactc
     2401 taggagcaag aaattgctaa agagggagat tttcgagctc aatatttaga catttcccca
     2461 taaactataa caggggtcta gaaagtgcat tggtcattgc ctgttggttt attctctctt
     2521 cttctttcct ttcttctttt tcttttctct cccgccttct ttttatgtct ttggagacag
     2581 tctatgtagc ccaagttgac tcggactgcc cactaattag ttcaagctga cctcaaactc
     2641 aaagcagcgt tccccacccc ccagcctccc gagtgcaagg attatagaca tacaccactc
     2701 tgcccagctc atctggtcac ttttgtccaa agagaaacat ttagcagctc tttctgctct
     2761 tgtcctgaag cagcatagag gaaacaatac ttttgtttaa gatgcctgct gagttggaca
     2821 gatgaagcct gtaaccccag atattcagga aatcgaatca gaagactctc aagttcaagg
     2881 ctgagctggc aacttaatga gatcttaata agaaggcaca gggggtagga agctggggag
     2941 gacccaagtt tggaacccaa gaactcctgg agagtcattg gggaagacct ctgacattga
     3001 cctctgacct ccacacacag gatcactcgc accctcctgt acatgggcac acagccccat
     3061 ttatatgtta cacactcccg cagcatgcac acaaattgct gctaatactg ggagagaagg
     3121 agccacaaac cccagggaaa actggatttc ccgtctgctt gctgaggaag agatgtggaa
     3181 caccattgat aaacacaaaa ctggactggg attaaatatt tgtgtccagt atcaggatcc
     3241 actagtgaat tcgaagttcc tatactccta taagaatagg aacttcggaa taggaacttc
     3301 atgatggaag ttcatatact ttctagagaa taggaacttc ggaataggaa cttcataact
     3361 tcgtatagga tactttatac gaagttatca gctgccgctc gaggcgcgtg atcttacggc
     3421 attatacgta tgatcggtcc acgatcagct agattatcta gtcagcttga tgtcatagct
     3481 gtttcctgag gctcaatact gaccatttaa atcatacctg acctccatag cagaaagtca
     3541 aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca actttcacca
     3601 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     3661 ggaagctaaa atgagccata ttcaacggga aacgtcttgc ttgaagccgc gattaaattc
     3721 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3781 tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3841 caaaggtagc gttgccaatg atgttacaga tgagatggtc aggctaaact ggctgacgga
     3901 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3961 caccactgcg atcccaggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     4021 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     4081 taattgtcct tttaacggcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     4141 taacggtttg gttggtgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     4201 agtctggaaa gaaatgcata aactcttgcc attctcaccg gattcagtcg tcactcatgg
     4261 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     4321 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     4381 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     4441 tatgaataaa ttgcagtttc acttgatgct cgatgagttt ttctaatgag gacctaaatg
     4501 taatcacctg gctcaccttc gggtgggcct ttctgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg atgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct cggaaaaaga
     5041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat tttctaccga
     5161 agaaaggccc a
//
