LOCUS       40924_20201        4512 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-IRES, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4512)
  AUTHORS   Singla V, Romaguera-Ros M, Garcia-Verdugo JM, Reiter JF.
  TITLE     Ofd1, a human disease gene, regulates the length and distal 
            structure of centrioles
  JOURNAL   Dev. Cell 18 (3), 410-424 (2010)
  PUBMED    20230748
REFERENCE   2  (bases 1 to 4512)
  AUTHORS   Singla V, Hunkapiller J, Norman AR, Skarnes WC, Reiter JF.
  TITLE     The Floxin system: an efficient method for creating diverse targeted
            modifications in gene trap embryonic stem cell lines
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4512)
  AUTHORS   Singla V, Reiter JF, Skarnes WC.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JUL-2008) Biochemistry, University of California, San 
            Francisco, 1550 4th Street, Rock Hall 446, San Francisco, CA 94158, 
            USA
REFERENCE   4  (bases 1 to 4512)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4512)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev. Cell";
            date: "2010"; volume: "18"; issue: "3"; pages: "410-424"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (22-JUL-2008) Biochemistry, University of California, San Francisco,
            1550 4th Street, Rock Hall 446, San Francisco, CA 94158, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..525
                     /label=b-actin
                     /note="b-actin"
                     /regulatory_class="promoter"
     regulatory      520..529
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(532..565)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          566..831
                     /note="En2"
     misc_feature    839..858
                     /label=tandem stop codons
                     /note="tandem stop codons"
     misc_feature    879..1453
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1459..1490
                     /label=multiple cloning site
                     /note="multiple cloning site"
     polyA_signal    1664..1798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1816..1863
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1887..1903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1911..1927)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1935..1965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1980..2001)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2289..2877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3051..3908)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3909..4013)
                     /label=AmpR promoter
     primer_bind     4487..4503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttcctt ctgcaggagc gtacagaacc cagggccctg gcacccgtgc agaccctggc
       61 ccaccccacc tgggcgctca gtgcccaaga gatgtccaca cctaggatgt cccgcggtgg
      121 gtggggggcc cgagagacgg gcaggccggg ggcaggcctg gccatgcggg gccgaaccgg
      181 gcactgccca gcgtggggcg cgggggccac ggcgcgcgcc cccagccccc gggcccagca
      241 ccccaaggcg gccaacgcca aaactctccc tcctcctctt cctcaatctc gctctcgctc
      301 tttttttttt tcgcaaaagg aggggagagg gggtaaaaaa atgctgcact gtgcggcgaa
      361 gccggtgagt gagcggcgcg gggccaatca gcgtgcgccg ttccgaaagt tgccttttat
      421 ggctcgagcg gccgcggcgg cgccctataa aacccagcgg cgcgacgcgc caccaccgcc
      481 gagaccgcgt ccgccccgcg agcacagagc ctcgcctttg ccaccatggc tataacttcg
      541 tatagcatac attatacgaa cggtagtgat aggcctttta gctacatctg ccaatccatc
      601 tcattttcac acacacacac accactttcc ttctggtcag tgggcacatg tccagcctca
      661 agtttatatc accaccccca atgcccaaca cttgtatggc cttgggcggg tcatcccccc
      721 ccccaccccc agtatctgca acctcaagct agcttgggtg cgttggttgt ggataagtag
      781 ctagactcca gcaaccagta acctctgccc tttctcctcc atgacaacca ggtaccggta
      841 gtgactaata gctgatagaa ttccgccccc cccccccccc cctctccctc ccccccccct
      901 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
      961 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     1021 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     1081 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     1141 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     1201 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     1261 gaaagagtca aatggctctc ctaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1321 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1381 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1441 cacgatgata ataggatcgc atgcggcgcg ccactagtga tatcgctagc ggatccacca
     1501 cctacagccc agtggggtcg ctctagagcg acctcgaggg ggggctagat aagtaatgat
     1561 cataatcagc catatcacat ctgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1621 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctagatccg
     1801 ggggtaccgc gtcgagaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     1861 ttcgtcgacg cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2701 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4021 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4081 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4141 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4201 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     4261 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4321 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     4381 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     4441 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     4501 agtgccaagc tt
//
