LOCUS Exported 9132 bp ds-DNA circular SYN 28-JAN-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pCW57.1-DUX4_mal103 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9132) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Jun 29, 2017 from SnapGene Viewer 2.4.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9132 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 41..158 /note="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" promoter 212..526 /note="tight TRE promoter" /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 220..238 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 256..274 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 291..309 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 327..345 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 363..381 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 398..416 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 434..452 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" CDS 585..1859 /codon_start=1 /note="DUX4" /translation="MALPTPSDSTLPAEARGRGRRRRLVWTPSQSEALRACFERNPYPG IATRERLAQAIGIPEPRVQIWFQNERSRQLRQHRRESRPWPGRRGPPEGRRKRTAVTGS QTALLLRAFEKDRFPGIAAREELARETGLPESRIQIWFQNRRARHPGQGGRAPAQAGGL CSAAPGGGHPAPSWVAFAHTGAWGTGLPAPHVPCAPGALPQGAFVSQAARAAPALQPSQ AAPAEGISQPAPARGDFAYAAPAPPDGALSHPQAPRWPPHPGKSREDRDPQRDGLPGPC AVAQPGPAQAGPQGQGVLAPPTSQGSPWWGWGRGPQVAGAAWEPQAGAAPPPQPAPPDA SASARQGQMQGIPAPSQALQEPAPWSALPCGLLLDELLASPEFLQQAQPLLETEAPGEL EASEEAASLEAPLSEEEYRALLEEL" primer_bind complement(1886..1902) /note="KS primer" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1905..2029) /gene="mutant version of attR" /bound_moiety="LR Clonase(TM)" /note="attR2" /note="recombination site for the Gateway(R) LR reaction" CDS 2047..2064 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" promoter 2074..2584 /note="hPGK promoter" /note="human phosphoglycerate kinase 1 promoter" CDS 2594..3190 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /note="PuroR" /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" CDS 3191..3244 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /note="T2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 3245..3991 /codon_start=1 /product="improved tetracycline-controlled transactivator" /note="rtTA-Advanced" /note="In the Tet-On(R) system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline." /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" CDS complement(4449..4460) /codon_start=1 /product="Factor Xa recognition and cleavage site" /note="Factor Xa site" /translation="IEGR" LTR 4665..4898 /note="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4970..5091 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" rep_origin 5131..5266 /note="SV40 ori" /note="SV40 origin of replication" CDS 5338..6198 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6369..6957 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 7245..7266 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 7281..7311 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 7319..7335 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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 7343..7359 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 7380..7398 /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" promoter 7424..7652 /note="RSV promoter" /note="Rous sarcoma virus enhancer/promoter" LTR 7653..7833 /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7880..8005 /note="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8498..8731 /note="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." ORIGIN 1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga 61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca 181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac 241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag 301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt 361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt 421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg 481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta 541 gaactagtgg atcccccggg ctgcaggaat tcgcccttgc caccatggcc ctcccgacac 601 cctcggacag caccctcccc gcggaagccc ggggacgagg acggcgacgg agactcgttt 661 ggaccccgag ccaaagcgag gccctgcgag cctgctttga gcggaacccg tacccgggca 721 tcgccaccag agaacggctg gcccaggcca tcggcattcc ggagcccagg gtccagattt 781 ggtttcagaa tgagaggtca cgccagctga ggcagcaccg gcgggaatct cggccctggc 841 ccgggagacg cggcccgcca gaaggccggc gaaagcggac cgccgtcacc ggatcccaga 901 ccgccctgct cctccgagcc tttgagaagg atcgctttcc aggcatcgcc gcccgggagg 961 agctggccag agagacgggc ctcccggagt ccaggattca gatctggttt cagaatcgaa 1021 gggccaggca cccgggacag ggtggcaggg cgcccgcgca ggcaggcggc ctgtgcagcg 1081 cggcccccgg cgggggtcac cctgctccct cgtgggtcgc cttcgcccac accggcgcgt 1141 ggggaacggg gcttcccgca ccccacgtgc cctgcgcgcc tggggctctc ccacaggggg 1201 ctttcgtgag ccaggcagcg agggccgccc ccgcgctgca gcccagccag gccgcgccgg 1261 cagaggggat ctcccaacct gccccggcgc gcggggattt cgcctacgcc gccccggctc 1321 ctccggacgg ggcgctctcc caccctcagg ctcctcgctg gcctccgcac ccgggcaaaa 1381 gccgggagga ccgggacccg cagcgcgacg gcctgccggg cccctgcgcg gtggcacagc 1441 ctgggcccgc tcaagcgggg ccgcagggcc aaggggtgct tgcgccaccc acgtcccagg 1501 ggagtccgtg gtggggctgg ggccggggtc cccaggtcgc cggggcggcg tgggaacccc 1561 aagccggggc agctccacct ccccagcccg cgcccccgga cgcctccgcc tccgcgcggc 1621 aggggcagat gcaaggcatc ccggcgccct cccaggcgct ccaggagccg gcgccctggt 1681 ctgcactccc ctgcggcctg ctgctggatg agctcctggc gagcccggag tttctgcagc 1741 aggcgcaacc tctcctagaa acggaggccc cgggggagct ggaggcctcg gaagaggccg 1801 cctcgctgga agcacccctc agcgaggaag aataccgggc tctgctggag gagctttaga 1861 agggcgaatt cgatatcaag cttatcgata ccgtcgacct cgaccatagt gactggatat 1921 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga 1981 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt tagtaatgaa 2041 ccggtccacc accaccacca ccactaagga tccggggttg gggttgcgcc ttttccaagg 2101 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg 2161 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc 2221 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc 2281 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct 2341 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa 2401 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc 2461 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa 2521 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc 2581 ccagcaattc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt 2641 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac 2701 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg 2761 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg 2821 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc 2881 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca 2941 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg 3001 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt 3061 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt 3121 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa 3181 gcccggtgcc gaaggtagag gttctctcct cacttgtggt gatgttgaag aaaaccctgg 3241 tccaatgtct agactggaca agagcaaagt cataaacgga gctctggaat tactcaatgg 3301 tgtcggtatc gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc 3361 taccctgtac tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat 3421 gctggacagg catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct 3481 gcggaacaac gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt 3541 gcatctcggc acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt 3601 cctgtgtcag caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca 3661 ctttacactg ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga 3721 gacacctacc accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg 3781 gcagggagcc gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa 3841 acagctaaag tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat 3901 gctcccagcc gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga 3961 cgattttgac cttgacatgc tccccgggta actaagtaag gatcgatcca agatatcgta 4021 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 4081 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 4141 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 4201 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 4261 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 4321 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt 4381 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 4441 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 4501 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 4561 ccccgcctgt ttcgcctcgg cgtccggact agaggtacct ttaagaccaa tgacttacaa 4621 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 4681 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 4741 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 4801 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 4861 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 4921 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 4981 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5041 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5101 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 5161 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 5221 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctttc aaatatgtat 5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 6181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6481 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 7021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 7081 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 7141 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 7201 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 7261 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 7321 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca 7381 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa 7441 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga 7501 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa 7561 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata 7621 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag 7681 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 7741 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 7801 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc 7861 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc 7921 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa 7981 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga 8041 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc 8101 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg 8161 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc 8221 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac 8281 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca 8341 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat 8401 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga 8461 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct 8521 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac 8581 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac 8641 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg 8701 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca 8761 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt 8821 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac 8881 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat 8941 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa 9001 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt 9061 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa 9121 ccccgagggg ac //