LOCUS       sequence-216320-        6137 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses aa28 to C-terminus of Flp recombinase fused to GAI CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6137)
  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 6137)
  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: pBW2176_pCAG-PV1-GAI-L1-FlpO-28C-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..6137
                     /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             2183..2203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2213..2437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2677..2696)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2742..2797
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2796..2815)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3158..3174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3158..3174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3171..3193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3182..3198
                     /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(3206..3212,3213..3230,3231..3236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3251..3272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3357..3491
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3511..3645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3602..3621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3713..3730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3884..4472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3964..3983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4643..5434,5435..5503))
                     /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     5266..5285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5504..5608)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5639..6018
                     /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 actccggagg
      961 gtctggctcc ggatcaagtg gtggcagcgg taccggcgag aagatcgcca gctgtgccgc
     1021 cgagctgacc tacctgtgct ggatgatcac ccacaacggc accgccatca agagggccac
     1081 cttcatgagc tacaacacca tcatcagcaa cagcctgagc ttcgacatcg tgaacaagag
     1141 cctgcagttc aagtacaaga cccagaaggc caccatcctg gaggccagcc tgaagaagct
     1201 gatccccgcc tgggagttca ccatcatccc ttacaacggc cagaagcacc agagcgacat
     1261 caccgacatc gtgtccagcc tgcagctgca gttcgagagc agcgaggagg ccgacaaggg
     1321 caacagccac agcaagaaga tgctgaaggc cctgctgtcc gagggcgaga gcatctggga
     1381 gatcaccgag aagatcctga acagcttcga gtacaccagc aggttcacca agaccaagac
     1441 cctgtaccag ttcctgttcc tggccacatt catcaactgc ggcaggttca gcgacatcaa
     1501 gaacgtggac cccaagagct tcaagctggt gcagaacaag tacctgggcg tgatcattca
     1561 gtgcctggtg accgagacca agacaagcgt gtccaggcac atctactttt tcagcgccag
     1621 aggcaggatc gaccccctgg tgtacctgga cgagttcctg aggaacagcg agcccgtgct
     1681 gaagagagtg aacaggaccg gcaacagcag cagcaacaag caggagtacc agctgctgaa
     1741 ggacaacctg gtgcgcagct acaacaaggc cctgaagaag aacgccccct accccatctt
     1801 cgctatcaag aacggcccta agagccacat cggcaggcac ctgatgacca gctttctgag
     1861 catgaagggc ctgaccgagc tgacaaacgt ggtgggcaac tggagcgaca agagggcctc
     1921 cgccgtggcc aggaccacct acacccacca gatcaccgcc atccccgacc actacttcgc
     1981 cctggtgtcc aggtactacg cctacgaccc catcagcaag gagatgatcg ccctgaagga
     2041 cgagaccaac cccatcgagg agtggcagca catcgagcag ctgaagggca gcgccgaggg
     2101 cagcatcaga taccccgcct ggaacggcat catcagccag gaggtgctgg actacctgag
     2161 cagctacatc aacaggcgga tccccaagaa aaagcggaag gtgtgagaat tcctgtgcct
     2221 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2281 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2341 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2401 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2461 tggggctcta gggggtatcc ccacgcgccc tgtagcgtgt accgctggga gttcgtagac
     2521 ggaaacaaac gcagaatcca agcgcggccg cccaggatac atagattacc acaactccga
     2581 gcccttccac cgctagcgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     2641 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     2701 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     2761 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2821 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     2881 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     2941 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     3001 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     3061 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     3121 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3181 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3241 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3301 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     3361 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3421 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3481 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     3541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3601 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     3661 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4321 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4921 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga
     5641 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     5701 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     5761 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5821 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5881 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5941 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     6001 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     6061 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     6121 ggggcggggg ggggggg
//
