LOCUS       sequence-77897-e        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of the site-specific recombinase Dre and BFP under the
            control of the hGFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  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 7232)
  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: hGFAP-NLS-HA-Dre-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..7232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2298..2318
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2319..2345
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2352..3374
                     /product="site-specific recombinase"
                     /label=Dre recombinase
                     /note="Dre is a site-specific recombinase from 
                     bacteriophage D6. Recombination occurs at rox sequences."
     CDS             3384..3440
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3441..4139
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(4159..4176)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4165..4389
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4564..4581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4735..5323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4815..4834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5494..6285,6286..6354))
                     /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"
     promoter        complement(6355..6459)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6486..6941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6573..6592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6783..6804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7072..7120
                     /note="synthetic polyadenylation signal"
     misc_feature    7134..7225
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7174..7193
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgagctccca cctccctctc
       61 tgtgctggga ctcacagagg gagacctcag gaggcagtct gtccatcaca tgtccaaatg
      121 cagagcatac cctgggctgg gcgcagtggc gcacaactgt aattccagca ctttgggagg
      181 ctgatgtgga aggatcactt gagcccagaa gttctagacc agcctgggca acatggcaag
      241 accctatctc tacaaaaaaa gttaaaaaat cagccacgtg tggtgacaca cacctgtagt
      301 cccagctatt caggaggctg aggtgagggg atcacttaag gctgggaggt tgaggctgca
      361 gtgagtcgtg gttgcgccac tgcactccag cctgggcaac agtgagaccc tgtctcaaaa
      421 gacaaaaaaa aaaaaaaaaa aaaaaagaac atatcctggt gtggagtagg ggacgctgct
      481 ctgacagagg ctcgggggcc tgagctggct ctgtgagctg gggaggaggc agacagccag
      541 gccttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc
      601 atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg
      661 cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc
      721 ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta
      781 ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt
      841 gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag
      901 gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggac tgggcagggt
      961 tctgaccctg tgggaccaga gtggagggcg tagatggacc tgaagtctcc agggacaaca
     1021 gggcccaggt ctcaggctcc tagttgggcc cagtggctcc agcgtttcca aacccatcca
     1081 tccccagagg ttcttcccat ctctccaggc tgatgtgtgg gaactcgagg aaataaatct
     1141 ccagtgggag acggaggggt ggccagggaa acggggcgct gcaggaataa agacgagcca
     1201 gcacagccag ctcatgtgta acggctttgt ggagctgtca aggcctggtc tctgggagag
     1261 aggcacaggg aggccagaca aggaaggggt gacctggagg gacagatcca ggggctaaag
     1321 tcctgataag gcaagagagt gccggccccc tcttgcccta tcaggacctc cactgccaca
     1381 tagaggccat gattgaccct tagacaaagg gctggtgtcc aatcccagcc cccagcccca
     1441 gaactccagg gaatgaatgg gcagagagca ggaatgtggg acatctgtgt tcaagggaag
     1501 gactccagga gtctgctggg aatgaggcct agtaggaaat gaggtggccc ttgagggtac
     1561 agaacaggtt cattcttcgc caaattccca gcaccttgca ggcacttaca gctgagtgag
     1621 ataatgcctg ggttatgaaa tcaaaaagtt ggaaagcagg tcagaggtca tctggtacag
     1681 cccttccttc cctttttttt tttttttttt gtgagacaag gtctctctct gttgcccagg
     1741 ctggagtggc gcaaacacag ctcactgcag cctcaaccta ctgggctcaa gcaatcctcc
     1801 agcctcagcc tcccaaagtg ctgggattac aagcatgagc caccccactc agccctttcc
     1861 ttccttttta attgatgcat aataattgta agtattcatc atggtccaac caaccctttc
     1921 ttgacccacc ttcctagaga gagggtcctc ttgcttcagc ggtcagggcc ccagacccat
     1981 ggtctggctc caggtaccac ctgcctcatg caggagttgg cgtgcccagg aagctctgcc
     2041 tctgggcaca gtgacctcag tggggtgagg ggagctctcc ccatagctgg gctgcggccc
     2101 aaccccaccc cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt
     2161 ctagcccact ccttcataaa gccctcgcat cccaggagcg agcagagcca gagcaggttg
     2221 gagaggagac gcatcacctc cgctgctcgc ggggatctgc gatctaagta agcttgaatt
     2281 ctcgagccgc caccatgcct aagaagaaga ggaaggttta cccatacgat gttccagatt
     2341 acgctggatc ctctgagctg attattagtg gttcatctgg tggattcctg cgaaacatcg
     2401 gcaaagagta tcaggaggcc gctgaaaact tcatgaggtt tatgaatgac cagggggcgt
     2461 acgctcctaa cactttgagg gatttgaggt tggtctttca tagctgggcc agatggtgcc
     2521 atgctcggca gcttgcatgg tttccaatta gtcctgaaat ggcacgcgaa tactttcttc
     2581 agttgcacga tgcagacctg gcctccacta ccatcgacaa gcactatgct atgcttaata
     2641 tgcttctgtc ccactgcgga ctgccaccct tgtccgacga caagtcagtg agtcttgcca
     2701 tgagaagaat tagaagagaa gccgcaaccg aaaagggtga gaggacagga caggcaatcc
     2761 ccctgcgctg ggacgacctg aagctgctgg atgtgctgct cagcaggagc gagcggctgg
     2821 tcgacctgcg caacagggct ttcctgttcg tagcctataa caccctcatg agaatgtctg
     2881 aaatatcacg catcagggtt ggggacttgg atcagacagg agacacagtg accctgcaca
     2941 tcagtcacac taagacaatc accacagctg cgggccttga caaagtgctc tcccggcgaa
     3001 ccacagcagt gctcaatgac tggctggacg tcagtgggct tagagaacat ccagacgctg
     3061 tgctcttccc acctatacac cggtcaaaca aagcccgcat tactaccacg cccctgaccg
     3121 cccctgccat ggagaagatt ttcagtgatg cctgggtgct gctgaacaaa cgggacgcca
     3181 cccccaataa agggaggtat aggacctgga ccggccattc cgccagggtg ggtgccgcaa
     3241 tagacatggc cgagaaacag gtgtctatgg tcgagattat gcaggaaggg acatggaaga
     3301 agcctgaaac actgatgcgg tatctcagaa ggggcggagt gtccgtggga gccaattctc
     3361 gactgatgga tagcggcagc ggcgccacga acttctctct gttaaagcaa gcaggcgacg
     3421 tggaagaaaa ccccggtccc agcgagctga ttaaggagaa catgcacatg aagctgtaca
     3481 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
     3541 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     3601 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     3661 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     3721 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     3781 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     3841 agaaaacact cggctgggag gccttcaccg agaccctgta ccccgctgac ggcggcctgg
     3901 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca
     3961 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     4021 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
     4081 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag ctgaattagg
     4141 tcagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4201 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4261 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4321 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4381 gctctatggc ttctgaggcg gaaagaacca gctggggaga tccgtcgacc gatgcccttg
     4441 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     4501 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
     4561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4921 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5161 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6481 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6541 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6601 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6661 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6721 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6781 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6841 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6901 atttaacgcg aattttaaca aaatattaac gtttacaatt tcccattcgc cattcaggct
     6961 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
     7021 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
     7081 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
     7141 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
     7201 agtgcaggtg ccagaacatt tctctatcga ta
//
