LOCUS       Exported File           8003 bp ds-DNA    circular SYN 29-SEP-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pSBtet_BP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8003)
  AUTHORS   Eric Kowarz
  TITLE     .
  JOURNAL   .
FEATURES             Location/Qualifiers
     source          1..8003
                     /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..5389
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5407..5704
                     /note="bGH-pA"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    5711..5757
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    5717..5758
                     /note="5'-proxITR"
                     /note="modified region proximal to 5'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    5759..5983
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
     misc_feature    5984..5987
                     /note="tata"
                     /note="color: #cc0000; direction: RIGHT"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6930..7790)
                     /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: 6930 .. 7721 / #ccffcc
                        2: 7722 .. 7790 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7791..7895)
                     /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 tgaccgagta
     4801 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4861 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4921 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4981 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5041 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5101 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5161 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5221 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5281 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5341 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat tcgaaggcct
     5401 gtcgtgaagc ttggggatca attctctaga gctcgctgat cagcctcgac tgtgccttct
     5461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5701 gcctactagc tactcgggac cccttaccga aacatcgccg cattctgcag aggagtcgag
     5761 tgtatgtaaa cttctgaccc actgggaatg tgatgaaaga aataaaagct gaaatgaatc
     5821 attctctcta ctattattct gatatttcac attcttaaaa taaagtggtg atcctaactg
     5881 acctaagaca gggaattttt actaggatta aatgtcagga attgtgaaaa agtgagttta
     5941 aatgtatttg gctaaggtgt atgtaaactt ccgacttcaa ctgtataggg atccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7921 gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     7981 agcgttaata ttttgttaaa att
//
