LOCUS       40924_20196        5274 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-IRES-eGFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  AUTHORS   Singla V, Hunkapiller J.
  TITLE     Efficient engineering of novel alleles at gene trap loci in mouse 
            embryonic stem cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5274)
  AUTHORS   Singla V, Hunkapiller J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUL-2009) Biochemistry, UCSF, 1550 4th Street, San 
            Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 5274)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5274)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUL-2009) Biochemistry, UCSF, 1550 4th Street, San Francisco, CA
            94158, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5274
                     /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    879..1453
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1506..2222
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    2426..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2578..2625
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(2649..2665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2673..2689)
                     /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(2697..2727)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2742..2763)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3813..4670)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4671..4775)
                     /label=AmpR promoter
     primer_bind     5249..5265
                     /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 tctgctagcg aagcttgccg
     1501 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1561 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1621 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1681 ccaccctcgt gaccaccttc acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1741 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1801 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1861 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1921 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1981 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2041 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2101 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2161 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2221 agtaaagcgg ccgctcgagc gccatcgcta gcggatccac cacctacagc ccagtggggt
     2281 cgctctagag cgacctcgag ggggggctag ataagtaatg atcataatca gccatatcac
     2341 atctgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     2401 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     2461 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2521 tttgtccaaa ctcatcaatg tatcttatca tgtctagatc cgggggtacc gcgtcgagaa
     2581 gttcctattc cgaagttcct attctctaga aagtatagga acttcgtcga cgcgaattcg
     2641 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2701 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2761 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2821 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2881 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2941 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3481 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4741 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4801 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     4861 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4921 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4981 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5041 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     5101 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5161 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5221 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctt
//
