LOCUS       Exported File           7238 bp ds-DNA    circular SYN 29-SEP-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pSBtet_pur
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  AUTHORS   Eric Kowarz
  TITLE     .
  JOURNAL   .
FEATURES             Location/Qualifiers
     source          1..7238
                     /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..3961
                     /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: 3218 .. 3835 / #993366 / rTetR
                        2: 3836 .. 3841 / #cc99b2
                        3: 3842 .. 3877 / #993366 / VP16 minimal transcriptio...
                        4: 3878 .. 3880 / #cc99b2
                        5: 3881 .. 3916 / #993366 / VP16 minimal transcriptio...
                        6: 3917 .. 3919 / #cc99b2
                        7: 3920 .. 3955 / #993366 / VP16 minimal transcriptio...
                        8: 3956 .. 3961 / #cc99b2"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             3959..4024
                     /codon_start=1
                     /note="P2A"
                     /note="color: #cc0000"
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     CDS             4025..4624
                     /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    4642..4939
                     /note="bGH-pA"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    4946..4992
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    4952..4993
                     /note="5'-proxITR"
                     /note="modified region proximal to 5'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    4994..5218
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
     misc_feature    5219..5222
                     /note="tata"
                     /note="color: #cc0000; direction: RIGHT"
     rep_origin      complement(5406..5994)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6165..7025)
                     /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: 6165 .. 6956 / #ccffcc
                        2: 6957 .. 7025 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7026..7130)
                     /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 tctagactgg acaagagcaa
     3241 agtcataaac ggcgctctgg aattactcaa tggagtcggt atcgaaggcc tgacgacaag
     3301 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     3361 gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg
     3421 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cattccgctg
     3481 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     3541 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     3601 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     3661 acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     3721 acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt
     3781 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     3841 ggccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt
     3901 tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg
     3961 gtccggcgct actaacttca gcctgctgaa gcaggctggt gacgtcgagg agaatcctgg
     4021 tcccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4081 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4141 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4201 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4261 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4321 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4381 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     4441 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     4501 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     4561 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     4621 ctgattcgaa ggcctgtcgt gaagcttggg gatcaattct ctagagctcg ctgatcagcc
     4681 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4741 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4801 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4861 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4921 gaaagaacca gctgggccta ctagctactc gggacccctt accgaaacat cgccgcattc
     4981 tgcagaggag tcgagtgtat gtaaacttct gacccactgg gaatgtgatg aaagaaataa
     5041 aagctgaaat gaatcattct ctctactatt attctgatat ttcacattct taaaataaag
     5101 tggtgatcct aactgaccta agacagggaa tttttactag gattaaatgt caggaattgt
     5161 gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta aacttccgac ttcaactgta
     5221 tagggatccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5281 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5341 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5821 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6061 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6241 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6301 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6361 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6421 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6481 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6541 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6601 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6661 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6721 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6781 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6841 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6901 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6961 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7021 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7081 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7141 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7201 catcaggaaa ttgtaagcgt taatattttg ttaaaatt
//
