LOCUS Exported File 7802 bp ds-DNA circular SYN 29-SEP-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pSBtet_BB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7802) AUTHORS Eric Kowarz TITLE . JOURNAL . FEATURES Location/Qualifiers source 1..7802 /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..5188 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /note="BSD" /note="confers resistance to blasticidin" /note="color: #ccffcc" /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 5206..5503 /note="bGH-pA" /note="bovine growth hormone polyadenylation signal" /note="color: #cc0000; direction: RIGHT" misc_feature 5510..5556 /note="3'-proxITR" /note="modified region proximal to 3'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 5516..5557 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 5558..5782 /note="3'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 5783..5786 /note="tata" /note="color: #cc0000; direction: RIGHT" rep_origin complement(5970..6558) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(6729..7589) /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: 6729 .. 7520 / #ccffcc    2: 7521 .. 7589 / #ccffcc / signal sequence" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7590..7694) /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 tggccatgcc 4801 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc 4861 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac 4921 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac 4981 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag 5041 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat 5101 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct 5161 gccctctggt tatgtgtggg agggctaatt cgaaggcctg tcgtgaagct tggggatcaa 5221 ttctctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 5281 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 5341 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 5401 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 5461 gtgggctcta tggcttctga ggcggaaaga accagctggg cctactagct actcgggacc 5521 ccttaccgaa acatcgccgc attctgcaga ggagtcgagt gtatgtaaac ttctgaccca 5581 ctgggaatgt gatgaaagaa ataaaagctg aaatgaatca ttctctctac tattattctg 5641 atatttcaca ttcttaaaat aaagtggtga tcctaactga cctaagacag ggaattttta 5701 ctaggattaa atgtcaggaa ttgtgaaaaa gtgagtttaa atgtatttgg ctaaggtgta 5761 tgtaaacttc cgacttcaac tgtataggga tccgcttcct cgctcactga ctcgctgcgc 5821 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5881 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5941 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 6001 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 6061 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 6121 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 6181 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 6241 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 6301 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 6361 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 6421 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 6481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 6541 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 6601 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 6661 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6721 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6781 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6841 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6901 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6961 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 7021 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 7081 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 7141 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 7201 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 7261 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 7321 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 7381 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 7441 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 7501 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 7561 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 7621 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 7681 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca 7741 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa 7801 tt //