LOCUS       sequence-217930-        4926 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into KpnI/EcoRI to fuse to GAI tag on 
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4926)
  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 4926)
  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: pBW2067_pCAG-PV1-GAI-L1-CTD-ENTRY-BGHpA
FEATURES             Location/Qualifiers
     source          1..4926
                     /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"
     polyA_signal    1009..1233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1473..1492)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1538..1593
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1592..1611)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1954..1970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1954..1970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1967..1989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1978..1994
                     /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(2002..2008,2009..2026,2027..2032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2047..2068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2153..2287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2307..2441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2344..2363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2398..2417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2509..2526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3439..4230,4231..4299))
                     /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     4062..4081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4300..4404)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4435..4814
                     /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 taccaagtct acgaattcct gtgccttcta
     1021 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1081 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1141 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1201 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1261 gctctagggg gtatccccac gcgccctgta gcgtgtaccg ctgggagttc gtagacggaa
     1321 acaaacgcag aatccaagcg cggccgccca ggatacatag attaccacaa ctccgagccc
     1381 ttccaccgct agcgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     1441 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     1501 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     1561 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     1621 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     1681 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     1741 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     1801 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     1861 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     1921 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1981 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2041 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2101 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     2161 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2221 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     2281 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     2341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2401 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     2461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt
     4441 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4501 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4561 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4621 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4681 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4741 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4801 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     4861 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     4921 cggggg
//
