LOCUS       sequence-386475-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component of the Bxb1-rcCre binary site-specific recombinase system,
            expression of the site-specific recombinase Bxb1 (mouse 
            codon-optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  AUTHORS   Hermann M, Stillhard P, Wildner H, Seruggia D, Kapp V, 
            Sanchez-Iranzo H, Mercader N, Montoliu L, Zeilhofer HU, Pelczar P
  TITLE     Binary recombinase systems for high-resolution conditional 
            mutagenesis.
  JOURNAL   Nucleic Acids Res. 2014 Jan 9.
  PUBMED    24413561
REFERENCE   2  (bases 1 to 6386)
  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: "Nucleic 
            Acids Res. 2014 Jan 9."
COMMENT     Sequence Label: pCAG-NLS-HA-Bxb1
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1689..1708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1689..1707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1727..1747
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1748..1774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3300..3317)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3306..3530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3602..3619)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3602..3618)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3611..3633)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3759..4214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4108..4127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4241..4345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4346..4414,4415..5206)
                     /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     complement(4564..4583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5377..5965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5866..5885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6119..6136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6253..6274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6289..6294,6295..6312,6313..6319)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6327..6343
                     /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)."
     primer_bind     6332..6354
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6351..6367
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6351..6367
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg ggnsgsgcgc crggsggggs ggggsggggs grggggsggg
      541 gsggggsgrg gsggrgrggk gsggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggk ggcggcaggk gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggrg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagatcc
     1681 ttaattaata atacgactca ctataggggg tcgacccgcc accatgccaa aaaagaaaag
     1741 aaaagtgtat ccctatgatg tccccgatta tgccggttca agagccctgg tcgtgattag
     1801 actgagccga gtgacagacg ccaccacaag tcccgagaga cagctggaat catgccagca
     1861 gctctgtgct cagcggggtt gggatgtggt cggcgtggca gaggatctgg acgtgagcgg
     1921 ggccgtcgat ccattcgaca gaaagaggag gcccaacctg gcaagatggc tcgctttcga
     1981 ggaacagccc tttgatgtga tcgtcgccta cagagtggac cggctgaccc gctcaattcg
     2041 acatctccag cagctggtgc attgggctga ggaccacaag aaactggtgg tcagcgcaac
     2101 agaagcccac ttcgatacta ccacaccttt tgccgctgtg gtcatcgcac tgatgggcac
     2161 tgtggcccag atggagctcg aagctatcaa ggagcgaaac aggagcgcag cccatttcaa
     2221 tattagggcc ggtaaataca gaggctccct gcccccttgg ggatatctcc ctaccagggt
     2281 ggatggggag tggagactgg tgccagaccc cgtccagaga gagcggattc tggaagtgta
     2341 ccacagagtg gtcgataacc acgaaccact ccatctggtg gcacacgacc tgaatagacg
     2401 cggcgtgctc tctccaaagg attattttgc tcagctgcag ggaagagagc cacagggaag
     2461 agaatggagt gctactgcac tgaagagatc tatgatcagt gaggctatgc tgggttacgc
     2521 aacactcaat ggcaaaactg tccgggacga tgacggagcc cctctggtga gggctgagcc
     2581 tattctcacc agagagcagc tcgaagctct gcgggcagaa ctggtcaaga ctagtcgcgc
     2641 caaacctgcc gtgagcaccc caagcctgct cctgagggtg ctgttctgcg ccgtctgtgg
     2701 agagccagca tacaagtttg ccggcggagg gcgcaaacat ccccgctatc gatgcaggag
     2761 catggggttc cctaagcact gtggaaacgg gacagtggcc atggctgagt gggacgcctt
     2821 ttgcgaggaa caggtgctgg atctcctggg tgacgctgag cggctggaaa aagtgtgggt
     2881 ggcaggatct gactccgctg tggagctggc agaagtcaat gccgagctcg tggatctgac
     2941 ttccctcatc ggatctcctg catatagagc tgggtcccca cagagagaag ctctggacgc
     3001 acgaattgct gcactcgctg ctagacagga ggaactggag ggcctggagg ccaggccctc
     3061 tggatgggag tggcgagaaa ccggacagag gtttggggat tggtggaggg agcaggacac
     3121 cgcagccaag aacacatggc tgagatccat gaatgtccgg ctcacattcg acgtgcgcgg
     3181 tggcctgact cgaaccatcg attttggcga cctgcaggag tatgaacagc acctgagact
     3241 ggggtccgtg gtcgaaagac tgcacactgg gatgtcctag gtcagagctc gctgatcagc
     3301 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3361 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3421 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3481 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3541 ggaaagaacc agctggggct cgagatccac tagttctagc ctcgaggcta gagcggccgc
     3601 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3661 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3721 gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt
     3781 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3841 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3901 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3961 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4021 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4081 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4141 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4201 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6061 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6121 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6181 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6241 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6301 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6361 ctatgaccat gaggcgcgcc ggattc
//
