LOCUS       sequence-375045-        6419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insert protein fragment into MluI/BspEI to fuse to GID1/NLS tags on 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  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 6419)
  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..6419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..1143
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1067..1089
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1151..1170
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2354..2374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2384..2608
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2848..2867)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2913..2968
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2967..2986)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3329..3345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3329..3345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3342..3364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3353..3369
                     /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(3377..3383,3384..3401,3402..3407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3422..3443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3528..3662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3682..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3773..3792
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3884..3901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4055..4643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4135..4154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4814..5605,5606..5674))
                     /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     5437..5456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5675..5779)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5810..6189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6342..6393
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     6344..6364
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      301 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      361 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      421 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
      481 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
      541 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
      601 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
      661 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
      721 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
      781 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
      841 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      901 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      961 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1021 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1081 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1141 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgatt
     1201 aattcgagcg aaggcgcgcc cattactcgc atccattctc aggctgtctc gtctcgtctc
     1261 gcacgcgtaa gtctactccg gagggtctgg ctccggatca agtggtggca gcggtaccat
     1321 ggccgccagc gacgaagtga acctgatcga gagcagaacc gtggtgcccc tgaacacctg
     1381 ggtgctgatc tccaacttca aggtggccta caacatcctg cggaggcccg acggcacctt
     1441 caacagacac ctggccgagt acctggaccg gaaagtgacc gccaacgcca accctgtgga
     1501 cggcgtgttc agcttcgacg tgctgatcga ccggcggatc aacctgctga gccgggtgta
     1561 cagacccgcc tacgccgatc aggaacagcc cccctctatc ctggatctgg aaaagcccgt
     1621 ggatggcgac atcgtgcccg tgatcctgtt cttccacggc ggcagctttg cccacagcag
     1681 cgccaatagc gccatctacg acaccctgtg cagacggctc gtgggcctgt gcaaatgcgt
     1741 ggtggtgtcc gtgaactacc gcagagcccc cgagaaccct tacccctgcg cctacgatga
     1801 tggctggatc gccctgaact gggtcaacag cagaagctgg ctgaagtcca agaaagacag
     1861 caaggtgcac atctttctgg ccggcgatag cagcggcggc aatatcgccc ataacgtggc
     1921 cctgagagcc ggcgagtctg gcatcgatgt gctgggcaat atcctgctga accccatgtt
     1981 cggcggcaac gagcggaccg agagcgagaa gtctctggac ggcaagtact tcgtgaccgt
     2041 gcgggaccgg gactggtact ggaaggcctt tctgcccgag ggcgaggaca gagagcaccc
     2101 cgcctgcaat cccttcagcc ccagaggcaa aagcctggaa ggcgtgtcct tcccaaagtc
     2161 cctggtggtg gtggccggcc tggacctgat cagagattgg cagctggcct atgccgaggg
     2221 cctgaagaaa gccggccagg aagtgaagct gatgcacctg gaaaaggcca ccgtgggctt
     2281 ctacctgctg cccaacaaca accacttcca caacgtgatg gacgagatca gcgccttcgt
     2341 gaacgccgag tgccccaaga aaaagcggaa ggtgtgagaa ttcctgtgcc ttctagttgc
     2401 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2461 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2521 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2581 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2641 agggggtatc cccacgcgcc ctgtagcgtg taccgctggg agttcgtaga cggaaacaaa
     2701 cgcagaatcc aagcgcggcc gcccaggata catagattac cacaactccg agcccttcca
     2761 ccgctagcgc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     2821 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     2881 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     2941 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3001 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     3061 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     3121 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     3181 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     3241 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     3301 cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3481 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3541 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3601 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3661 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     3721 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3781 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4801 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4861 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4921 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4981 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5041 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5101 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5161 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5221 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5281 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5341 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5401 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5461 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5521 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5581 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5641 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5701 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5761 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg acattgatta
     5821 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5881 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5941 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     6001 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     6061 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     6121 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     6181 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     6241 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     6301 grrngggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     6361 cgtgtacggt gggaggtcta tataagcaga gctskytggc taactagaga acccactgc
//
