LOCUS Exported File 8216 bp ds-DNA circular SYN 29-SEP-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pSBtet_GN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8216) AUTHORS Eric Kowarz TITLE . JOURNAL . FEATURES Location/Qualifiers source 1..8216 /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..5602 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin)" /note="color: #ccffcc" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 5620..5917 /note="bGH-pA" /note="bovine growth hormone polyadenylation signal" /note="color: #cc0000; direction: RIGHT" misc_feature 5924..5970 /note="3'-proxITR" /note="modified region proximal to 3'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 5930..5971 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 5972..6196 /note="3'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 6197..6200 /note="tata" /note="color: #cc0000; direction: RIGHT" rep_origin complement(6384..6972) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(7143..8003) /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: 7143 .. 7934 / #ccffcc    2: 7935 .. 8003 / #ccffcc / signal sequence" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8004..8108) /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 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 4861 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 4921 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 4981 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 5041 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 5101 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 5161 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 5221 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 5281 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 5341 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 5401 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 5461 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 5521 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 5581 ccttcttgac gagttcttct gattcgaagg cctgtcgtga agcttgggga tcaattctct 5641 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 5701 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 5761 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 5821 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc 5881 tctatggctt ctgaggcgga aagaaccagc tgggcctact agctactcgg gaccccttac 5941 cgaaacatcg ccgcattctg cagaggagtc gagtgtatgt aaacttctga cccactggga 6001 atgtgatgaa agaaataaaa gctgaaatga atcattctct ctactattat tctgatattt 6061 cacattctta aaataaagtg gtgatcctaa ctgacctaag acagggaatt tttactagga 6121 ttaaatgtca ggaattgtga aaaagtgagt ttaaatgtat ttggctaagg tgtatgtaaa 6181 cttccgactt caactgtata gggatccgct tcctcgctca ctgactcgct gcgctcggtc 6241 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6301 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6361 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6421 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6481 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6541 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6601 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6661 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 6721 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 6781 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 6841 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6901 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6961 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7021 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7261 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 7741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 7801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 7861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8101 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg 8161 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaatt //