LOCUS       40924_20206        4529 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-MCS2-IRES-MCS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4529)
  AUTHORS   Singla VI, Julie H, Santos N, Seol A, Norman AR, Wakenight P, 
            Skarnes WC, Reiter JF.
  TITLE     A resource for engineering alleles in mouse embryonic stem cells
  JOURNAL   Nat. Methods (2010) In press
REFERENCE   2  (bases 1 to 4529)
  AUTHORS   Reiter JF.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2009) Biochemistry, UCSF, 1550 4th Street, Rock 
            Hall 446, San Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 4529)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4529)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods (2010) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-NOV-2009) Biochemistry, UCSF, 1550 4th Street, Rock Hall 446, 
            San Francisco, CA 94158, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..525
                     /label=b-actin promoter
                     /note="b-actin promoter"
                     /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..826
                     /note="Engrailed2 intron"
     misc_feature    827..873
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    874..892
                     /label=tandem stops
                     /note="tandem stops"
     misc_feature    902..1467
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1476..1507
                     /label=multiple cloning site
                     /note="multiple cloning site"
     polyA_signal    1681..1815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1833..1880
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1904..1920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1928..1944)
                     /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(1952..1982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1997..2018)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2306..2894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3068..3925)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3926..4030)
                     /label=AmpR promoter
     primer_bind     4504..4520
                     /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 ggtaccggag
      841 ctcagcgcta gatctatcga tacgcgtacc ggtagtgact aatagctgat agaattccgc
      901 ccccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
      961 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     1021 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     1081 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     1141 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     1201 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     1261 tgagttggat agttgtggaa agagtcaaat ggctctccta agcgtattca acaaggggct
     1321 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     1381 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     1441 ggttttcctt tgaaaaacac gatgataata ggatcgcatg cggcgcgcca ctagtgatat
     1501 cgctagcgga tccaccacct acagcccagt ggggtcgctc tagagcgacc tcgagggggg
     1561 gctagataag taatgatcat aatcagccat atcacatctg tagaggtttt acttgcttta
     1621 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1681 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1741 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1801 tatcatgtct agatccgggg gtaccgcgtc gagaagttcc tattccgaag ttcctattct
     1861 ctagaaagta taggaacttc gtcgacgcga attcgtaatc atggtcatag ctgtttcctg
     1921 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1981 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2101 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     2761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3061 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3241 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3481 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3721 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3901 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3961 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4021 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4081 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     4141 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4201 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4261 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     4321 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     4381 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4441 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4501 gttgtaaaac gacggccagt gccaagctt
//
