LOCUS       Exported                5662 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses N-terminus to aa27 of Flp recombinase fused to FRB CID.
ACCESSION   .
VERSION     .
KEYWORDS    pBW2272_pCAG-PV1-FlpO-N27-L1-FRB-NLS-BGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5662)
  AUTHORS   Weinberg BH, Cho JH, Agarwal Y, Pham NTH, Caraballo LD, Walkosz M, 
            Ortega C, Trexler M, Tague N, Law B, Benman WKJ, Letendre J, Beal J 
            , Wong WW
  TITLE     High-performance chemical- and light-inducible recombinases in 
            mammalian cells and mice
  JOURNAL   Nature Communications (2019) volume 10, Article number: 4845
REFERENCE   2  (bases 1 to 5662)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5662
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(22..38)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(22..38)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(35..57)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    46..62
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(70..100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    115..136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      221..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    375..509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(412..431)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     466..485
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(577..594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(748..1336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(828..847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1507..2367)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2130..2149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2368..2472)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2503..2882
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2884..3159
                     /label=chicken beta-actin promoter
     intron          3160..4175
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4099..4121
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4183..4202
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4433..4708
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             4709..4729
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4739..4963
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5203..5222)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5268..5323
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5322..5341)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
       61 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      181 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      481 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
      541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2461 aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta
     2521 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2581 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2641 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2701 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2761 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2821 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2881 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     2941 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
     3001 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggaaaggt
     3061 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     3121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg
     3181 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
     3241 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
     3301 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
     3361 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     3421 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
     3481 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
     3541 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     3601 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
     3661 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     3721 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     3781 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     3841 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     3901 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     3961 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     4021 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     4081 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     4141 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     4201 tgctgtctca tcattttggc aaagaattga ttaattcgag cgaaggcgcg cccattactc
     4261 gcatccattc tcaggctgtc tcgtctcgtc tcgcacgcgt gccgccacca tgagccagtt
     4321 cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga
     4381 gaggcccagc tccggagggt ctggctccgg atcaagtggt ggcagcggta ccatcctctg
     4441 gcatgagatg tggcatgaag gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa
     4501 cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct atgatggaac ggggccccca
     4561 gactctgaag gaaacatcct ttaatcaggc ctatggtcga gatttaatgg aggcccaaga
     4621 gtggtgcagg aagtacatga aatcagggaa tgtcaaggac ctcctccaag cctgggacct
     4681 ctattatcat gtgttccgac gaatctcacc caagaaaaag cggaaggtgt gagaattcct
     4741 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4801 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4861 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4921 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4981 accagctggg gctctagggg gtatccccac gcgccctgta gcgtgtaccg ctgggagttc
     5041 gtagacggaa acaaacgcag aatccaagcg cggccgccca ggatacatag attaccacaa
     5101 ctccgagccc ttccaccgct agcgcactcc tcaggtgcag gctgcctatc agaaggtggt
     5161 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     5221 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5281 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5341 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5401 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5461 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5521 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     5581 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     5641 atggagatcc ctcgacctgc ag
//
