LOCUS       sequence-218066-        7017 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses aa28 to aa396 of Flp recombinase fused to GAI and ABI 
            CIDs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7017)
  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   Nat Commun. 2019 Oct 24;10(1):4845. doi: 10.1038/s41467-019-12800-7.
  PUBMED    31649244
REFERENCE   2  (bases 1 to 7017)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-24"; volume: "10(1):4845. doi"; pages: " 
            10.1038/s41467-019-12800-7"
COMMENT     Sequence Label: pBW2614_pCAG-PV1-GAI-L2-28-396-FlpO-ABI-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..7017
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             689..706
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3041..3064
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3065..3085
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3095..3319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3559..3578)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3624..3679
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3678..3697)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4040..4056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4040..4056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4053..4075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4064..4080
                     /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(join(4088..4094,4095..4112,4113..4118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4133..4154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4239..4373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4393..4527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4430..4449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4484..4503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4595..4612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4766..5354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4846..4865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5525..6316,6317..6385))
                     /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     6148..6167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6386..6490)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6521..6900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga aggcgcgccc attactcgca tccattctca ggctgtctcg tctcgtctcg
      661 cacgcgtgcc gccaccatga agcgggacca ccaccatcac catcatcagg acaagaaaac
      721 catgatgatg aacgaagagg acgacggcaa cggcatggac gagctgctgg ctgtgctggg
      781 ctacaaagtg cggagcagcg agatggccga cgtggcccag aaactggaac agctggaagt
      841 gatgatgagc aacgtgcagg aagatgacct gtcccagctg gccaccgaga cagtgcacta
      901 caaccccgcc gagctgtaca cctggctgga ctccatgctg accgacctga acgggccggc
      961 cggcggcggt agcggcggtg gctctggcgg tcctgcaggg ggcgagaaga tcgccagctg
     1021 tgccgccgag ctgacctacc tgtgctggat gatcacccac aacggcaccg ccatcaagag
     1081 ggccaccttc atgagctaca acaccatcat cagcaacagc ctgagcttcg acatcgtgaa
     1141 caagagcctg cagttcaagt acaagaccca gaaggccacc atcctggagg ccagcctgaa
     1201 gaagctgatc cccgcctggg agttcaccat catcccttac aacggccaga agcaccagag
     1261 cgacatcacc gacatcgtgt ccagcctgca gctgcagttc gagagcagcg aggaggccga
     1321 caagggcaac agccacagca agaagatgct gaaggccctg ctgtccgagg gcgagagcat
     1381 ctgggagatc accgagaaga tcctgaacag cttcgagtac accagcaggt tcaccaagac
     1441 caagaccctg taccagttcc tgttcctggc cacattcatc aactgcggca ggttcagcga
     1501 catcaagaac gtggacccca agagcttcaa gctggtgcag aacaagtacc tgggcgtgat
     1561 cattcagtgc ctggtgaccg agaccaagac aagcgtgtcc aggcacatct actttttcag
     1621 cgccagaggc aggatcgacc ccctggtgta cctggacgag ttcctgagga acagcgagcc
     1681 cgtgctgaag agagtgaaca ggaccggcaa cagcagcagc aacaagcagg agtaccagct
     1741 gctgaaggac aacctggtgc gcagctacaa caaggccctg aagaagaacg ccccctaccc
     1801 catcttcgct atcaagaacg gccctaagag ccacatcggc aggcacctga tgaccagctt
     1861 tctgagcatg aagggcctga ccgagctgac aaacgtggtg ggcaactgga gcgacaagag
     1921 ggcctccgcc gtggccagga ccacctacac ccaccagatc accgccatcc ccgaccacta
     1981 cttcgccctg gtgtccaggt actacgccta cgaccccatc agcaaggaga tgatcgccct
     2041 gaaggacgag accaacccca tcgaggagtg gcagcacatc gagcagctga agggcagcgc
     2101 cgagggctcc ggagggtctg gctccggatc aagtggtggc agcggtaccc ctttgtatgg
     2161 ttttacttcg atttgtggaa gaagacctga gatggaagat gctgtttcga ctataccaag
     2221 attccttcaa tcttcctctg gttcgatgtt agatggtcgg tttgatcctc aatccgccgc
     2281 tcatttcttc ggtgtttacg acggccatgg cggttctcag gtagcgaact attgtagaga
     2341 gaggatgcat ttggctttgg cggaggagat agctaaggag aaaccgatgc tctgcgatgg
     2401 tgatacgtgg ctggagaagt ggaagaaagc tcttttcaac tcgttcctga gagttgactc
     2461 ggagattggg tcagttgcgc cggagacggt tgggtcaacg tcggtggttg ccgttgtttt
     2521 cccgtctcac atcttcgtcg ctaactgcgg tgactctaga gccgttcttt gccgcggcaa
     2581 aactgcactt ccattatccg ttgaccataa accggataga gaagatgaag ctgcgaggat
     2641 tgaagccgca ggagggaaag tgattcagtg gaatggagct cgtgttttcg gtgttctcgc
     2701 catgtcgaga tccattggcg atagatactt gaaaccatcc atcattcctg atccggaagt
     2761 gacggctgtg aagagagtaa aagaagatga ttgtctgatt ttggcgagtg acggggtttg
     2821 ggatgtaatg acggatgaag aagcgtgtga gatggcaagg aagcggattc tcttgtggca
     2881 caagaaaaac gcggtggctg gggatgcatc gttgctcgcg gatgagcgga gaaaggaagg
     2941 gaaagatcct gcggcgatgt ccgcggctga gtatttgtca aagctggcga tacagagagg
     3001 aagcaaagac aacataagtg tggtggtggt tgatttgaag gattacaagg acgatgacga
     3061 taagcccaag aaaaagcgga aggtgtgaga attcctgtgc cttctagttg ccagccatct
     3121 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3181 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3241 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3301 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3361 ccccacgcgc cctgtagcgt gtaccgctgg gagttcgtag acggaaacaa acgcagaatc
     3421 caagcgcggc cgcccaggat acatagatta ccacaactcc gagcccttcc accgctagcg
     3481 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     3541 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     3601 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     3661 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     3721 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     3781 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     3841 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     3901 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     3961 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     4021 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4201 gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccag
     4261 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4321 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4381 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4501 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     4561 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4621 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4681 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4741 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4801 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4861 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4921 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4981 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5041 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5101 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5161 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5221 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5281 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5341 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5401 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5461 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5521 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5641 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5701 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5821 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5941 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6181 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6361 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6421 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6481 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactagt
     6541 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6601 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6661 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6721 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6781 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6841 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6901 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     6961 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg ggggggg
//
