LOCUS Exported File 8429 bp ds-DNA circular SYN 29-SEP-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pSBtet_BH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8429) AUTHORS Eric Kowarz TITLE . JOURNAL . FEATURES Location/Qualifiers source 1..8429 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 8..11 /note="tata" /note="color: #cc0000" misc_feature 8..11 /note="tata" /note="color: #cc0000" misc_feature 12..236 /note="5'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 242..283 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 288..669 /note="TCE-Promoter" /note="Tet-responsive promoter" /note="color: #cc0000; direction: RIGHT" misc_feature 670..682 /note="5'-SfiI" /note="GGCCTCTGAGGCC" /note="color: #a6acb3" CDS 686..2338 /codon_start=1 /note="luc2" /note="firefly luciferase" /note="color: #cc0000" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" misc_feature 2339..2351 /note="3'-SfiI" /note="GGCCTGTCAGGCC" /note="color: #a6acb3" polyA_signal 2366..2586 /note="SV40-polyA" /note="SV40 polyadenylation signal" /note="color: #cc0000; direction: RIGHT" promoter 2593..3204 /note="RPBSA" /note="constitutive promoter" /note="color: #cc0000; direction: RIGHT" promoter 2593..3027 /note="RPL13a promoter" /note="color: #ffffff; direction: RIGHT" exon 3027..3079 /note="RPL41 exon1" /note="color: #cc0000; direction: RIGHT" intron 3080..3194 /note="RPL41 intron1" /note="color: #cc0000; direction: RIGHT" exon 3195..3204 /note="RPL41 exon2" /note="color: #cc0000; direction: RIGHT" CDS 3218..3916 /codon_start=1 /note="mTag-BFP" /note="color: #cc0000" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" CDS 3917..3982 /codon_start=1 /note="P2A" /note="color: #cc0000" /translation="GSGATNFSLLKQAGDVEENPGP" CDS 3983..4726 /codon_start=1 /product="improved tetracycline-controlled transactivator" /note="rtTA-Advanced" /note="binds to a Tet-responsive element in the presence but not the absence of doxycycline" /note="This feature has 8 segments:    1: 3983 .. 4600 / #993366 / rTetR    2: 4601 .. 4606 / #cc99b2    3: 4607 .. 4642 / #993366 / VP16 minimal transcriptio...    4: 4643 .. 4645 / #cc99b2    5: 4646 .. 4681 / #993366 / VP16 minimal transcriptio...    6: 4682 .. 4684 / #cc99b2    7: 4685 .. 4720 / #993366 / VP16 minimal transcriptio...    8: 4721 .. 4726 / #cc99b2" /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" CDS 4724..4789 /codon_start=1 /note="P2A" /note="color: #cc0000" /translation="GSGATNFSLLKQAGDVEENPGP" CDS 4790..5815 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="HygR" /note="confers resistance to hygromycin" /note="color: #ccffcc" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 5833..6130 /note="bGH-pA" /note="bovine growth hormone polyadenylation signal" /note="color: #cc0000; direction: RIGHT" misc_feature 6137..6183 /note="3'-proxITR" /note="modified region proximal to 3'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 6143..6184 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 6185..6409 /note="3'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 6410..6413 /note="tata" /note="color: #cc0000; direction: RIGHT" rep_origin complement(6597..7185) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(7356..8216) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments:    1: 7356 .. 8147 / #ccffcc    2: 8148 .. 8216 / #ccffcc / signal sequence" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8217..8321) /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: LEFT" ORIGIN 1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact 61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta 121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta 181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt 241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agacgagttt 301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg 361 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt 421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta 481 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta 541 tgtcgattag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca 601 gatcgcctgg agccaattcc aactcttttg ccttatacct actttccgta ccacttccta 661 ccctcgaaag gcctctgagg ccaccatgga agatgccaaa aacattaaga agggcccagc 721 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg 781 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac 841 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct 901 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt 961 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg 1021 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg 1081 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat 1141 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt 1201 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat 1261 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca 1321 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat 1381 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac 1441 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct 1501 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt 1561 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat 1621 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca 1681 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac 1741 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa 1801 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt 1861 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat 1921 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt 1981 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc 2041 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct 2101 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac 2161 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct 2221 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc 2281 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaagg 2341 cctgtcaggc caagcttcca tcgatagaca tgataagata cattgatgag tttggacaaa 2401 ccacaacaag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt 2461 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 2521 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat 2581 gtggtaccta ggcgagaccc tgtctcacaa aataaagtaa gcccggactg agtgcggaaa 2641 ggcgggcctg gcgggtctgg tctccccatg cgggccacca gaggccctgc agccttcagt 2701 cgcttgaagg ggtaatggcg cttccactca caaacatggc ggacagagcg tgtgaacgag 2761 atgaacagcc cctcaaaaat atggccgccg aggctggacg gccgtgcccc agcagcaccg 2821 cctccgcgcc ccacgtgatc tctcgccggg cacagcgctg accgcggagg tccaaccgga 2881 agaatgtccg gattggacat tcggaagagg gcccgccttc cctggggaat ctctgcgcac 2941 gcgcagaacg cttcgaccaa tgaaaacaca ggaagccgtc cgcgcaaccg cgttgcgtca 3001 cttctgccgc ccctgtttca aggtatatag ccgtagacgg aacttcgcct ttctctcggc 3061 cttagcgcca tttttttggg tgagtgtttt ttggttcctg cgttgggatt ccgtgtacaa 3121 tccatagaca tctgacctcg gcacttagca tcatcacagc aaactaactg tagcctttct 3181 ctctttccct gtagaaacct ctgcacctga ggccaccatg agcgagctga ttaaggagaa 3241 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc 3301 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg 3361 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac 3421 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt 3481 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac 3541 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc 3601 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta 3661 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag 3721 ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa 3781 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga 3841 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact 3901 ggggcacaag ctgaacggca gtggagctac taacttcagc ctgctgaagc aggctggtga 3961 cgtcgaggag aatcctggcc ccatgtctag actggacaag agcaaagtca taaacggcgc 4021 tctggaatta ctcaatggag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa 4081 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga 4141 tgccctgcca atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga 4201 gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca 4261 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct 4321 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc 4381 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt 4441 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc 4501 aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat 4561 catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg acgcccttga 4621 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct 4681 gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtccg gcgctactaa 4741 cttcagcctg ctgaagcagg ctggtgacgt cgaggagaat cctggtccca tgaaaaagcc 4801 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga 4861 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg 4921 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta 4981 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag 5041 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc 5101 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc 5161 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata 5221 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac 5281 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg 5341 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt 5401 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga 5461 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca 5521 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc 5581 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga 5641 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt 5701 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt 5761 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagttcga 5821 aggcctgtcg tgaagcttgg ggatcaattc tctagagctc gctgatcagc ctcgactgtg 5881 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 5941 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 6001 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 6061 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 6121 agctgggcct actagctact cgggacccct taccgaaaca tcgccgcatt ctgcagagga 6181 gtcgagtgta tgtaaacttc tgacccactg ggaatgtgat gaaagaaata aaagctgaaa 6241 tgaatcattc tctctactat tattctgata tttcacattc ttaaaataaa gtggtgatcc 6301 taactgacct aagacaggga atttttacta ggattaaatg tcaggaattg tgaaaaagtg 6361 agtttaaatg tatttggcta aggtgtatgt aaacttccga cttcaactgt atagggatcc 6421 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 6481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 6541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 6601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 6661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 6721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 6781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 6841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 6901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 6961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 7021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 7081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 7141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 7201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 7261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 7321 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 7381 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 7441 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 7501 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 7561 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 7621 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 7681 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 7741 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 7801 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 7861 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 7921 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 7981 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 8041 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 8101 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 8161 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 8221 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 8281 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 8341 ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa 8401 attgtaagcg ttaatatttt gttaaaatt //