LOCUS       sequence-217922-        5529 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into MluI/BspEI to fuse to GID1/NLS tags on 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5529)
  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 5529)
  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: pBW2066_pCAG-PV1-NTD-ENTRY-L1-GID1-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     289..311
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     373..392
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1576..1596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1606..1830
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2070..2089)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2135..2190
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2189..2208)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2551..2567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2551..2567)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2564..2586)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2575..2591
                     /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(2599..2605,2606..2623,2624..2629))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2644..2665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2750..2884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2904..3038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2941..2960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2995..3014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3106..3123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3277..3865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3357..3376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4036..4827,4828..4896))
                     /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     4659..4678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4897..5001)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5032..5411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
       61 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
      121 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
      181 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
      241 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
      301 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
      361 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattga
      421 ttaattcgag cgaaggcgcg cccattactc gcatccattc tcaggctgtc tcgtctcgtc
      481 tcgcacgcgt aagtctactc cggagggtct ggctccggat caagtggtgg cagcggtacc
      541 atggccgcca gcgacgaagt gaacctgatc gagagcagaa ccgtggtgcc cctgaacacc
      601 tgggtgctga tctccaactt caaggtggcc tacaacatcc tgcggaggcc cgacggcacc
      661 ttcaacagac acctggccga gtacctggac cggaaagtga ccgccaacgc caaccctgtg
      721 gacggcgtgt tcagcttcga cgtgctgatc gaccggcgga tcaacctgct gagccgggtg
      781 tacagacccg cctacgccga tcaggaacag cccccctcta tcctggatct ggaaaagccc
      841 gtggatggcg acatcgtgcc cgtgatcctg ttcttccacg gcggcagctt tgcccacagc
      901 agcgccaata gcgccatcta cgacaccctg tgcagacggc tcgtgggcct gtgcaaatgc
      961 gtggtggtgt ccgtgaacta ccgcagagcc cccgagaacc cttacccctg cgcctacgat
     1021 gatggctgga tcgccctgaa ctgggtcaac agcagaagct ggctgaagtc caagaaagac
     1081 agcaaggtgc acatctttct ggccggcgat agcagcggcg gcaatatcgc ccataacgtg
     1141 gccctgagag ccggcgagtc tggcatcgat gtgctgggca atatcctgct gaaccccatg
     1201 ttcggcggca acgagcggac cgagagcgag aagtctctgg acggcaagta cttcgtgacc
     1261 gtgcgggacc gggactggta ctggaaggcc tttctgcccg agggcgagga cagagagcac
     1321 cccgcctgca atcccttcag ccccagaggc aaaagcctgg aaggcgtgtc cttcccaaag
     1381 tccctggtgg tggtggccgg cctggacctg atcagagatt ggcagctggc ctatgccgag
     1441 ggcctgaaga aagccggcca ggaagtgaag ctgatgcacc tggaaaaggc caccgtgggc
     1501 ttctacctgc tgcccaacaa caaccacttc cacaacgtga tggacgagat cagcgccttc
     1561 gtgaacgccg agtgccccaa gaaaaagcgg aaggtgtgag aattcctgtg ccttctagtt
     1621 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1681 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1741 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1801 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     1861 ctagggggta tccccacgcg ccctgtagcg tgtaccgctg ggagttcgta gacggaaaca
     1921 aacgcagaat ccaagcgcgg ccgcccagga tacatagatt accacaactc cgagcccttc
     1981 caccgctagc gcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     2041 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     2101 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     2161 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     2221 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     2281 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     2341 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     2401 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     2461 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     2521 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2581 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2641 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2701 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     2761 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2821 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2881 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     2941 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3001 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     3061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
     5041 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5101 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5161 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5221 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5281 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5341 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5401 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     5461 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     5521 ggggggggg
//
