LOCUS       Exported File           8021 bp ds-DNA    circular SYN 29-SEP-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pSBtet_GP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Eric Kowarz
  TITLE     .
  JOURNAL   .
FEATURES             Location/Qualifiers
     source          1..8021
                     /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..5407
                     /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    5425..5722
                     /note="bGH-pA"
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    5729..5775
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    5735..5776
                     /note="5'-proxITR"
                     /note="modified region proximal to 5'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    5777..6001
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
     misc_feature    6002..6005
                     /note="tata"
                     /note="color: #cc0000; direction: RIGHT"
     rep_origin      complement(6189..6777)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6948..7808)
                     /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: 6948 .. 7739 / #ccffcc
                        2: 7740 .. 7808 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7809..7913)
                     /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 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     4861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     5161 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     5281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5401 tgcctgattc gaaggcctgt cgtgaagctt ggggatcaat tctctagagc tcgctgatca
     5461 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     5521 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     5581 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     5641 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     5701 gcggaaagaa ccagctgggc ctactagcta ctcgggaccc cttaccgaaa catcgccgca
     5761 ttctgcagag gagtcgagtg tatgtaaact tctgacccac tgggaatgtg atgaaagaaa
     5821 taaaagctga aatgaatcat tctctctact attattctga tatttcacat tcttaaaata
     5881 aagtggtgat cctaactgac ctaagacagg gaatttttac taggattaaa tgtcaggaat
     5941 tgtgaaaaag tgagtttaaa tgtatttggc taaggtgtat gtaaacttcc gacttcaact
     6001 gtatagggat ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6061 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7921 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7981 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat t
//
