LOCUS       sequence-227095-        5574 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into MluI/BspEI to fuse to ABI/NLS tags on 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5574)
  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 5574)
  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: pBW1312_pCAG-PV1-NTD-ENTRY-L1-ABI-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..5574
                     /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             1609..1632
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1633..1653
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1663..1887
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2127..2146)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2192..2247
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2246..2265)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2608..2624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2608..2624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2621..2643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2632..2648
                     /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(2656..2662,2663..2680,2681..2686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2701..2722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2807..2941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2961..3095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2998..3017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3052..3071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3163..3180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3334..3922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3414..3433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4093..4884,4885..4953))
                     /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     4716..4735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4954..5058)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5089..5468
                     /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 cacgcgtaag tctactccgg agggtctggc tccggatcaa gtggtggcag cggtacccct
      721 ttgtatggtt ttacttcgat ttgtggaaga agacctgaga tggaagatgc tgtttcgact
      781 ataccaagat tccttcaatc ttcctctggt tcgatgttag atggtcggtt tgatcctcaa
      841 tccgccgctc atttcttcgg tgtttacgac ggccatggcg gttctcaggt agcgaactat
      901 tgtagagaga ggatgcattt ggctttggcg gaggagatag ctaaggagaa accgatgctc
      961 tgcgatggtg atacgtggct ggagaagtgg aagaaagctc ttttcaactc gttcctgaga
     1021 gttgactcgg agattgggtc agttgcgccg gagacggttg ggtcaacgtc ggtggttgcc
     1081 gttgttttcc cgtctcacat cttcgtcgct aactgcggtg actctagagc cgttctttgc
     1141 cgcggcaaaa ctgcacttcc attatccgtt gaccataaac cggatagaga agatgaagct
     1201 gcgaggattg aagccgcagg agggaaagtg attcagtgga atggagctcg tgttttcggt
     1261 gttctcgcca tgtcgagatc cattggcgat agatacttga aaccatccat cattcctgat
     1321 ccggaagtga cggctgtgaa gagagtaaaa gaagatgatt gtctgatttt ggcgagtgac
     1381 ggggtttggg atgtaatgac ggatgaagaa gcgtgtgaga tggcaaggaa gcggattctc
     1441 ttgtggcaca agaaaaacgc ggtggctggg gatgcatcgt tgctcgcgga tgagcggaga
     1501 aaggaaggga aagatcctgc ggcgatgtcc gcggctgagt atttgtcaaa gctggcgata
     1561 cagagaggaa gcaaagacaa cataagtgtg gtggtggttg atttgaagga ttacaaggac
     1621 gatgacgata agcccaagaa aaagcggaag gtgtgagaat tcctgtgcct tctagttgcc
     1681 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1741 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1801 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1861 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     1921 gggggtatcc ccacgcgccc tgtagcgtgt accgctggga gttcgtagac ggaaacaaac
     1981 gcagaatcca agcgcggccg cccaggatac atagattacc acaactccga gcccttccac
     2041 cgctagcgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     2101 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     2161 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     2221 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     2281 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     2341 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     2401 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     2461 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     2521 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     2581 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2761 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2941 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     3001 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3061 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     3121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4261 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4621 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4921 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4981 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5041 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga cattgattat
     5101 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5161 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5221 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5281 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5341 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5401 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5461 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     5521 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg gggg
//
