LOCUS Exported 7232 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Co-expression of the site-specific recombinase Dre and BFP under the control of the hGFAP promoter. ACCESSION . VERSION . KEYWORDS hGFAP-NLS-HA-Dre-P2A-BFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7232 /organism="synthetic DNA construct" /mol_type="other DNA" 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 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(5494..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" primer_bind 6117..6136 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" 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 //