LOCUS       sequence-77891-e        6770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Component of the Co-Driver binary site-specific recombinase system, 
            generates functional rox60-71Cre upon removal of an internal STOP 
            cassette by Dre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6770)
  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 6770)
  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-Roxed-Cre
FEATURES             Location/Qualifiers
     source          1..6770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..670
                     /label=chicken beta-actin promoter
     intron          671..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1692..1711
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1692..1710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1730..1750
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1875..1906
                     /label=rox
                     /bound_moiety="Dre recombinase"
     polyA_signal    2094..2228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2348..2482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2602..2736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2765..2796
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             3649..3672
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3695..3712)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3701..3925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3997..4014)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3997..4013)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4006..4028)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4155..4610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4242..4261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4452..4473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4636..4740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4741..4809,4810..5601)
                     /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(4959..4978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5772..6360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6261..6280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6514..6531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6648..6669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6684..6689,6690..6707,6708..6714)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6722..6738
                     /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     6727..6749
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6746..6762
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6746..6762
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcgccggatt cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
      421 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
      481 ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
      541 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      601 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      661 gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg
      721 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      781 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct
      841 gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc gggggggagc ggctcggggg
      901 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg
      961 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc ggtcgggctg taaccccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtgcg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     1261 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct ggcggagccg aaatctggga ggcgccgccg
     1441 caccccctct agcgggcgcg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc atctccagcc tcggggctgc
     1561 cgcaggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacaga
     1681 tccttaatta ataatacgac tcactatagg gggtcgaccc gccaccatgc ccaagaagaa
     1741 gaggaaggtg acctctgatg aagtcaggaa gaacctgatg gacatgttca gggacaggca
     1801 ggccttctct gaacacacct ggaagatgct cctgtctgtg tgcagatcct gggctgcctg
     1861 gtgcaagctg aacgtaactt taaataattg gcattattta aagttaatgt atgcgctacg
     1921 aagttattcg cgatgaataa atgaaagctt gcagatctgc gactctagag gatctgcgac
     1981 tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2041 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2101 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2161 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     2221 tctggatctg cgactctaga ggatcataat cagccatacc acatttgtag aggttttact
     2281 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2341 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2461 tgtatcttat catgtctgga tctgcgactc tagaggatca taatcagcca taccacattt
     2521 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2581 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2641 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2701 tccaaactca tcaatgtatc ttatcatgtc tggatcccca tcaagctgat ccggaaccct
     2761 taattaactt taaataattg gcattattta aagttaaaca ggaaatggtt ccctgctgaa
     2821 cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc tgtgaagacc
     2881 atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct gcctcgccct
     2941 tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa tgtggatgct
     3001 ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca agtcagatcc
     3061 ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct gggcattgcc
     3121 tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga catctcccgc
     3181 accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt gtccacagct
     3241 ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg gatctctgtg
     3301 tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa gaatggtgtg
     3361 gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat ctttgaggcc
     3421 acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct ggcctggtct
     3481 ggccactctg ccagagtggg tgctgccagg gacatggcca gggctggtgt gtccatccct
     3541 gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta catcagaaac
     3601 ctggactctg agactggggc catggtgagg ctgctcgagg atggggacga ttacaaggat
     3661 gacgacgata agtaggtcag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3721 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3781 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3841 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3901 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgaga
     3961 tccactagtt ctagcctcga ggctagagcg gccgccactg gccgtcgttt tacaacgtcg
     4021 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4081 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4141 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4201 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4501 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4561 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4621 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4681 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4741 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4801 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4861 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4921 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4981 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5041 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5101 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5161 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5221 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5281 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5341 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5401 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5461 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5521 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5581 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5641 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5701 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5761 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5821 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5881 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5941 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6001 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6061 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6481 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6541 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6601 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6661 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6721 attgtgagcg gataacaatt tcacacagga aacagctatg accatgaggc
//
