LOCUS       sequence-77881-e        6588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the site-specific recombinase B3 (mouse 
            codon-optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  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 6588)
  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-B3
FEATURES             Location/Qualifiers
     source          1..6588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1683..1702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1683..1701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1721..1741
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1742..1768
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3504..3521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3510..3734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3806..3823)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3806..3822)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3815..3837)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3964..4419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4051..4070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4261..4282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4445..4549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4550..4618,4619..5410)
                     /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(4768..4787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5581..6169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6070..6089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6323..6340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6457..6478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6493..6498,6499..6516,6517..6523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6531..6547
                     /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     6536..6558
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6555..6571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6555..6571
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      481 gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag atccttaatt
     1681 aataatacga ctcactatag ggggtcgacc cgccaccatg cctaagaaga agaggaaggt
     1741 ttacccatac gatgttccag attacgctgg atccagcagc tatatggatc tggtggatga
     1801 cgagcccgca acactgtatc acaagtttgt cgaatgtctg aaggcaggag agaatttctg
     1861 cggcgacaag ctgtccggaa tcattacaat ggccatcctg aaagccatta aggctctgac
     1921 tgaggtgaag aaaaccacct tcaacaagta caaaactacc atcaagcagg ggctgcagta
     1981 tgatgtgggc agctccacca tttctttcgt ctaccacctg aaagactgtg atgagctgag
     2041 tcgcggcctg tcagacgctt ttgaacctta caagttcaaa atcaagagta ataaggaggc
     2101 aaccagcttc aagacactgt tcaggggccc cagcttcggc agccagaaga actggcgaaa
     2161 gaaagaggtg gatagagaag tggacaatct gttccatagc accgagacag atgaatccat
     2221 ctttaagttc attctgaaca cactggatag catcgagact cagaccaata cagaccgcca
     2281 gaaaacagtg ctgactttca ttctgctgat gaccttcttt aactgctgta ggaacaatga
     2341 tctgatgaat gtggacccca gcaccttcaa gatcgtgaaa aacaagttcg tcggctacct
     2401 gctgcaggct gaggtgaaac agactaagac ccgaaaaagc aggaacatct tctttttccc
     2461 tatcagagaa aaccggtttg atctgttcct ggcactgcac gactttttca gaacttgcca
     2521 gcctacccca aagtctcggc tgagtgacca ggtgagtgag cagaaatggc agctgtttcg
     2581 cgattcaatg gtcatcgact ataaccgctt tttccgaaag tttccagctt cccccatctt
     2641 cgcaattaag cacggaccta aatctcacct ggggaggcat ctgatgaaca gtttcctgca
     2701 taagaatgag ctggatagct gggccaactc cctggggaat tggtctagtt cacagaacca
     2761 gcgagaaagc ggagctcgac tgggatacac ccacggcgga agagacctgc cacagcctct
     2821 gtttggattc ctggccgggt attgtgtgcg gaatgaggaa ggccatatcg tcggactggg
     2881 gctggagaag gacatcaacg atctgtttga cggaattatg gaccccctga acgagaaaga
     2941 agacacagag atttgcgaat catacggaga atgggctaag atcgtgagca aagacgtcct
     3001 gatttttctg aagagatatc acagcaaaaa cgcctgcagg agataccaga acagcactct
     3061 gtatgctcgg acattcctga agactgagtc cgtgaccctg tcagggagca aaggctctga
     3121 ggaacctagc tccccagtca gaatccccat tctgagcatg ggcaaggcca gcccttctga
     3181 gggaaggaaa ctgagagcat ctgaacatgc caacgacgat aatgagatcg aaaagattga
     3241 ctcagattct agtcagagcg aggaaatccc aattgagatg agtgattcag aggacgaaac
     3301 aactgcatca aacatcagcg gcatctacct ggacatgtct aaggccaaca gtaatgtggt
     3361 ctattccccc ccttctcaga ctggaagagc agctggagca ggacgaaagc gaggagtggg
     3421 cggcaggagg accgtcgagt ctaaacgaag gagagtgctg gcccccatca accggggaag
     3481 ttgagtcaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3541 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3601 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3661 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3721 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgagat ccactagttc
     3781 tagcctcgag gctagagcgg ccgccactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3841 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3901 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3961 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4021 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4081 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4141 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4201 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4261 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4321 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4381 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4441 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4501 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5101 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5161 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5401 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5461 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5521 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5761 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5821 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5881 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5941 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6001 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6061 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6361 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6421 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6481 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6541 ataacaattt cacacaggaa acagctatga ccatgaggcg cgccggat
//
