LOCUS       40924_20191        7034 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFlipTrap, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  AUTHORS   Trinh Le A, Hochgreb T, Graham M, Wu D, Ruf-Zamojski F, Jayasena CS,
            Saxena A, Hawk R, Gonzalez-Serricchio A, Dixson A, Chow E, Gonzales 
            C, Leung HY, Solomon I, Bronner-Fraser M, Megason SG, Fraser SE.
  TITLE     A versatile gene trap to visualize and interrogate the function of 
            the vertebrate proteome
  JOURNAL   Genes Dev. 25 (21), 2306-2320 (2011)
  PUBMED    22056673
REFERENCE   2  (bases 1 to 7034)
  AUTHORS   Trinh LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-AUG-2011) Biology, California Institute of Technology,
            1200 E. California Blvd, 139-74, Pasadena, CA 91125, USA
REFERENCE   3  (bases 1 to 7034)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7034)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            Dev."; date: "2011"; volume: "25"; issue: "21"; pages: "2306-2320"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-AUG-2011) Biology, California Institute of Technology, 1200 E. 
            California Blvd, 139-74, Pasadena, CA 91125, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    548..581
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     intron          644..969
                     /gene="RalGDS/AF-6"
                     /note="splice acceptor"
     protein_bind    complement(1000..1033)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1039..1752
                     /label=Citrine
                     /note="enhanced variant of YFP (Heikal et al., 2001)"
     intron          1780..2104
                     /gene="RalGDS/AF-6"
                     /note="splice donor"
     protein_bind    complement(2111..2144)
                     /label=lox2272
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      complement(2151..2506)
                     /label=alpha tubulin
                     /note="alpha tubulin"
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(2516..3223)
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     protein_bind    3238..3271
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3278..3311
                     /label=lox2272
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_recomb     3318..3351
                     /label=FRT-F3
                     /note="FRT-F3"
     primer_bind     complement(3912..3928)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3965..3983)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4004..4020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4028..4044)
                     /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(4052..4082)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4097..4118)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4406..4994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5168..6025)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6026..6130)
                     /label=AmpR promoter
     rep_origin      complement(6156..6611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     6753..6769
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6779..6797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta cttagggaaa
       61 attttactca attaaaagta aaagtatctg gctagaatct tacttgagta aaagtaaaaa
      121 agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa
      181 actagagatt cttgtttaag cttttaatct caaaaaacat taaatgaaat gcatacaagg
      241 ttttatcctg ctttagaact gtttgtattt aattatcaaa ctataagaca gacaatctaa
      301 tgccagtaca cgctactcaa agttgtaaaa cctcagattt aacttcagta gaagctgatt
      361 ctcaaaattg ttagtgtcaa gcctagctct tttggggctg aaaagcaatc ctgcagtgct
      421 gaaaagcctc tcacaggcag ccgatgcggg aagaggtgta ttagtcttga tagagaggct
      481 gcaaatagca ggaaacgtga gcagagactc cctggtgtct gaaacacagg ccagatgggc
      541 cctcgaggaa gttcctattc tctagaaagt ataggaactt cactagtcta cgcccccaac
      601 tgagagaact caaaggttac cccagttggg gcactacatc gattcaggaa cctcacagac
      661 tgcaggttga gctccccctg aagggctcac taacaccctt tttttcacaa caaccaagtt
      721 tttttcatgt ggtcttccat ccaggtacta accaggctta gccctgctta gcttcagtga
      781 gtaaccagtc ttgggcagca agttgatttg gctgtaccaa taatgatttt tgtaataata
      841 atttttgcaa ttagaatgct actgattgag atattacaca ctccaatttg atctgaaact
      901 aaaagtttct gttgtcatta cagaggaaat tcctgactct tttgataatg taaatgtgct
      961 gtgttgcagg acgaactggt cgctacacat tcacccggga taacttcgta tagcatacat
     1021 tatacgaagt tatccggagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1081 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1141 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1201 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac
     1261 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1321 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1381 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1441 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1501 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1561 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1621 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     1681 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1741 gagctgtaca agaccggtcc ttcaaacatt gggctacagg tgagtgtacc tgaagaaaga
     1801 acacaattta cttacactta catttcaaac agataacttg acttagattg cttttagttt
     1861 tttggtgtgg actaagtcca tttgacattg aggttaattt aattggtgtg aaagttttcg
     1921 ccaaactcag gcacataaca gaaagctaaa aacccaattg ttatgaaagc ggctagtcct
     1981 aaatcaatga gtggattaag aaattcacac agaagaaacc tttttaaaac gcctagaaac
     2041 ctggtagcta ggtcatataa ctgagttcct gaggtgaaag ccccttattg taaactaccc
     2101 aaagggatcc ataacttcgt ataggatact ttatacgaag ttattacgta actaatctac
     2161 ttttccttca tcccaagtgg ttggaagcca aacctgccaa agattagtaa gcagattact
     2221 tatcttcaat gcccagaatg tcttaaaacc aaaagcttat gttaacatgt acaagaaaga
     2281 tttttaaaga actgcacatg gacagattgt agcacatcac catcaattta ttccaaattt
     2341 ggcaagcaaa ttccacaaat aataagcata ctggccataa caacacaaca actgatcaag
     2401 aatgtgtctc taacatttag gagtagactt tacacttgat acctcatgtt tatcgtgtgt
     2461 aaattagcgg tctactgtgt tcagttttag acatttagtg gatattgata tcttacttgt
     2521 acagctcgtc catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga
     2581 tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg
     2641 gcagctgcac gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc
     2701 cgtccttcag cttgagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca
     2761 tccgctcgga ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg
     2821 ggaagttggt gccgcgcagc ttcaccttgt agatgaactc gccgtcctgg agggaggagt
     2881 cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc
     2941 cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct
     3001 tggagccgta catgaactga ggggacagga tgtcccaggc gaagggcagg gggccaccct
     3061 tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct
     3121 cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg caccttgaag cgcatgaact
     3181 ccttgatgat ggccatgtta tcctcctcgc ccttgctcac cacggctccc gtcgacaata
     3241 acttcgtata atgtatgcta tacgaagtta tcatatgata acttcgtata aagtatccta
     3301 tacgaagtta tttcgaagaa gttcctattc ttcaaatagt ataggaactt cagatcaatt
     3361 cgcgaagata cggccacggg tgctcttgat cctgtggctg attttggact gtgctgctcg
     3421 cagctgctga tgaatcacat acttcctcca ttttcttcca ctgattgact gttataattt
     3481 ccctaatttc caggtcaagg tgctgtgcat tgtggtaata gatgtgacat gacgtcactt
     3541 ccaaaggacc aatgaacatg tctgaccaat ttcatataat gtgaaaacga ttttcatagg
     3601 cagaataaat aacatttaaa ttaaactggg catcagcgca attcaattgg tttggtaata
     3661 gcaagggaaa atagaatgaa gtgatctcca aaaaataagt actttttgac tgtaaataaa
     3721 attgtaagga gtaaaaagta cttttttttc taaaaaaatg taattaagta aaagtaaaag
     3781 tattgatttt taattgtact caagtaaagt aaaaatcccc aaaaataata cttaagtaca
     3841 gtaatcaagt aaaattactc aagtacttta cacctctggt tcttgacccc ctaccttcag
     3901 caagcccagc agatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt
     3961 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     4021 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4081 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4141 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4201 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4381 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5161 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5221 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5281 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5341 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5401 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5461 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5521 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5581 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5641 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5701 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5761 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5821 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5881 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5941 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6001 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6061 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6121 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     6181 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6241 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6301 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6361 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6421 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6481 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6541 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6601 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6661 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6721 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     6781 atacgactca ctatagggcg aattgggtac ccacaatgac ggctactaca tggtgccatt
     6841 ccttcctctt tataggaatg gagactacct cctgtccaac aaggctcttg gatacgagta
     6901 cgcctacctg ttggacccag gtcattgcac aacaccagaa atgccctctg atctgcaaaa
     6961 gacgtgaata tctgttcaga cacccatatc cactctgttc cacacaggtc agaggtttgt
     7021 ccaggagttc ttga
//
