LOCUS Exported File 8447 bp ds-DNA circular SYN 29-SEP-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pSBtet_GH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8447) AUTHORS Eric Kowarz TITLE . JOURNAL . FEATURES Location/Qualifiers source 1..8447 /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..3934 /codon_start=1 /product="enhanced GFP" /note="EGFP" /note="mammalian codon-optimized" /note="This feature has 3 segments:    1: 3218 .. 3220 / #05fd14    2: 3221 .. 3223 / #05fd14 / 1a    3: 3224 .. 3934 / #05fd14" /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3935..4000 /codon_start=1 /note="P2A" /note="color: #cc0000" /translation="GSGATNFSLLKQAGDVEENPGP" CDS 4001..4744 /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: 4001 .. 4618 / #993366 / rTetR    2: 4619 .. 4624 / #cc99b2    3: 4625 .. 4660 / #993366 / VP16 minimal transcriptio...    4: 4661 .. 4663 / #cc99b2    5: 4664 .. 4699 / #993366 / VP16 minimal transcriptio...    6: 4700 .. 4702 / #cc99b2    7: 4703 .. 4738 / #993366 / VP16 minimal transcriptio...    8: 4739 .. 4744 / #cc99b2" /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" CDS 4742..4807 /codon_start=1 /note="P2A" /note="color: #cc0000" /translation="GSGATNFSLLKQAGDVEENPGP" CDS 4808..5833 /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 5851..6148 /note="bGH-pA" /note="bovine growth hormone polyadenylation signal" /note="color: #cc0000; direction: RIGHT" misc_feature 6155..6201 /note="3'-proxITR" /note="modified region proximal to 3'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 6161..6202 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 6203..6427 /note="3'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 6428..6431 /note="tata" /note="color: #cc0000; direction: RIGHT" rep_origin complement(6615..7203) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(7374..8234) /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: 7374 .. 8165 / #ccffcc    2: 8166 .. 8234 / #ccffcc / signal sequence" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8235..8339) /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 ttgagcaagg gcgaggagct 3241 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt 3301 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat 3361 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg 3421 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc 3481 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa 3541 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 3601 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag 3661 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat 3721 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc 3781 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct 3841 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 3901 cgggatcact ctcggcatgg acgagctgta caagggcagt ggagctacta acttcagcct 3961 gctgaagcag gctggtgacg tcgaggagaa tcctggcccc atgtctagac tggacaagag 4021 caaagtcata aacggcgctc tggaattact caatggagtc ggtatcgaag gcctgacgac 4081 aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa 4141 caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt 4201 ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg 4261 ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga 4321 gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct 4381 ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga 4441 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc 4501 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct 4561 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg 4621 gccggccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga 4681 ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc 4741 cgggtccggc gctactaact tcagcctgct gaagcaggct ggtgacgtcg aggagaatcc 4801 tggtcccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa 4861 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag 4921 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta 4981 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct 5041 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt 5101 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc 5161 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc 5221 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca 5281 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct 5341 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga 5401 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag 5461 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg 5521 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg 5581 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt 5641 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg 5701 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac 5761 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag 5821 ggcaaaggaa tagttcgaag gcctgtcgtg aagcttgggg atcaattctc tagagctcgc 5881 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 5941 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 6001 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 6061 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 6121 tctgaggcgg aaagaaccag ctgggcctac tagctactcg ggacccctta ccgaaacatc 6181 gccgcattct gcagaggagt cgagtgtatg taaacttctg acccactggg aatgtgatga 6241 aagaaataaa agctgaaatg aatcattctc tctactatta ttctgatatt tcacattctt 6301 aaaataaagt ggtgatccta actgacctaa gacagggaat ttttactagg attaaatgtc 6361 aggaattgtg aaaaagtgag tttaaatgta tttggctaag gtgtatgtaa acttccgact 6421 tcaactgtat agggatccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 6481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 6541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 6601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 6661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 6721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 6781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 6841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 6901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 6961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 7021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 7081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 7141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 7201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 7261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 7321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 7381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 7441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 7501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 7561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 7621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 7681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 7741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 7801 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 7861 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 7921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 7981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 8041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 8101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 8161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 8221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 8281 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 8341 acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag 8401 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaatt //