LOCUS       sequence-77874-e        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component of the Bxb1-rcCre binary site-specific recombinase system,
            reverse complementary Cre CDS flanked by Bxb1 attB/P recognition 
            sites in head-to-head orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  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 5930)
  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-attBr-Cre-rc-attP
FEATURES             Location/Qualifiers
     source          1..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..673
                     /label=chicken beta-actin promoter
     intron          674..1682
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    1695..1744
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     CDS             complement(1757..1780)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2755..2775)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2791..2838
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     primer_bind     complement(2858..2875)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2864..3088
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3160..3177)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3160..3176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3169..3191)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3318..3773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3405..3424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3615..3636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3799..3903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3904..3972,3973..4764)
                     /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(4122..4141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4935..5523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5424..5443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5677..5694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5811..5832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5847..5852,5853..5870,5871..5877)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5885..5901
                     /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     5890..5912
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5909..5925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5909..5925
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggcgcgccgg attcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
      421 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
      481 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg
      541 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt gccccgctcc gcgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
      841 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
      901 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggcccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcgtgtgcg cgaggggagc
     1021 gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc ggcggtcggg ctgtaacccc
     1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1201 gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1261 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctggcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccatctcca gcctcggggc
     1561 tgccgcaggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agatccttaa ttaatcggcc ggcttgtcga cgacggcggt ctccgtcgtc aggatcatcc
     1741 gggctgacct gacctactta tcgtcgtcat ccttgtaatc tccccatcct cgagcagcct
     1801 caccatggcc ccagtctcag agtccaggtt tctgatgtag ttcatcacaa tgttcacatt
     1861 ggtccagcca ccagcctgca tgatttcagg gatggacaca ccagccctgg ccatgtccct
     1921 ggcagcaccc actctggcag agtggccaga ccaggccagg tatctctgcc cagagtcatc
     1981 cttggcacca tagatcaggc ggtgggtggc ctcaaagatc ccttccaggg cccgggtgga
     2041 cagttgggag gtggcagaag gggcagccac accattcttt ctgacccggc agaacaggta
     2101 gttgttgggg tcatcagcca caccagacac agagatccat ctctccacca gcttggtaac
     2161 ccccagggac agggccttct ccacaccagc tgtggacacc agggtcttgg tcctgccaat
     2221 gtggatcagc attctcccac catcggtgcg ggagatgtcc ttcactctga ttctggcaat
     2281 ttcggcaatg cgcagcaggg tgttgtaggc aatgcccagg aaggccaggt tcctgatgtc
     2341 ctggcatctg tcagagttct ccatcaggga tctgacttgg tcaaagtcag tgcgttcaaa
     2401 ggccagggcc tgcttggctc tctccccagc atccacattc tcctttctga ttctcctcat
     2461 caccagggac acagcattgg agtcagaagg gcgaggcagg ccagatctcc tgtgcagcat
     2521 gttgagctgg cccaggtgct gttggatggt cttcacagcc aggcctctgg cttgcaggta
     2581 caggaggtag tccctcacat cctcaggttc agcagggaac catttcctgt tgttcagctt
     2641 gcaccaggca gcccaggatc tgcacacaga caggagcatc ttccaggtgt gttcagagaa
     2701 ggcctgcctg tccctgaaca tgtccatcag gttcttcctg acttcatcag aggtcacctt
     2761 cctcttcttc ttgggcatgg tggtggcggc ggtttgtacc gtacaccact gagaccgcgg
     2821 tggttgacca gacaaaccac gagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2881 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2941 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3001 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3061 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg
     3121 agatccacta gttctagcct cgaggctaga gcggccgcca ctggccgtcg ttttacaacg
     3181 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3241 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3301 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3361 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3421 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3481 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3541 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3601 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3661 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3721 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3781 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3841 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3901 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3961 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4021 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4081 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4141 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4201 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4261 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4321 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4381 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4441 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4501 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4561 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4621 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4681 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4741 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4801 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4861 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4921 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5041 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5521 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5581 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5641 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5701 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5761 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5821 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5881 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
//
