LOCUS       Exported                6534 bp ds-DNA     circular SYN 05-SEP-2017
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 31, 2019 from SnapGene Viewer 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6534
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     207..240
                     /label=CMV-SgrDI-mut-f
                     /note="color: black; sequence: 
                     CGCGATGTACGGGCCAGATCGTCGACGTTGACATT"
     primer_bind     232..270
                     /label=CMV-D2i-SgrDI-f
                     /note="color: black; sequence: 
                     GATCGTCGACGGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     primer_bind     235..275
                     /label=SalI-CMV-D2ir-f
                     /note="color: black; sequence: 
                     TATGTCGACGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACG"
     TATA_signal     804..810
                     /note="color: #a6acb3"
     protein_bind    814..832
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     primer_bind     939..958
                     /label=TG1.4.9
                     /note="color: black; sequence: CAGTACggatccACCATGGGTTCTAG"
     CDS             948..1571
                     /label=hTetR
                     /note="color: #993366; direction: RIGHT"
     primer_bind     1570..1604
                     /label=TG1.4.7
                     /note="color: black; sequence: 
                     GGCCAAAAAAGAAGAGAAAGgctagcggcgccggc"
     primer_bind     complement(1570..1604)
                     /label=TG1.4.10
                     /note="color: black; sequence: 
                     gccggcgccgctagcCTTTCTCTTCTTTTTTGGCC"
     sig_peptide     1572..1589
                     /label=NLS
                     /note="color: #cc99b2; direction: RIGHT"
     gene            1588..1589
                     /gene="Med25VBD"
                     /label=Med25VBD
                     /note="color: #a6acb3; direction: RIGHT"
     gene            1608..2030
                     /gene="LOVpep"
                     /label=LOVpep
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1617..1619
                     /gene="T406"
                     /note="color: #a6acb3"
     misc_feature    1620..1622
                     /gene="T407"
                     /note="color: #a6acb3"
     misc_feature    1647..1649
                     /gene="V416"
                     /note="color: #a6acb3"
     misc_feature    1995..1997
                     /gene="I532A"
                     /note="color: #a6acb3"
     primer_bind     complement(2015..2051)
                     /label=TG1.4.11
                     /note="color: black; sequence: 
                     gtactgACCGGTttaGGAGCCGCCGCCGGAGGGGGCGGCGAAGGCGTAGGCGTTCCAGG
                     TCCAggcggcctcgtcgatg"
     primer_bind     complement(2029..2051)
                     /label=TG1.4.8
                     /note="color: black; sequence: 
                     gtactgACCGGTttagccgcggcggcggg"
     misc_feature    2031..2042
                     /gene="degron"
                     /note="color: #a6acb3"
     misc_feature    2076..2434
                     /label=WPRE
                     /note="color: #a6acb3; direction: RIGHT"
     polyA_signal    2492..2716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(2696..2716)
                     /label=bGH-pA-MluI-r
                     /note="color: black; sequence: 
                     ATATACGCGTCCATAGAGCCCACCGCATCCC"
     primer_bind     complement(2696..2716)
                     /label=bGH-pA-r
                     /note="color: black; sequence: CCATAGAGCCCACCGCATCCC"
     protein_bind    3000..3047
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
                     /note="color: #31849b; direction: RIGHT"
     CDS             3055..4077
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #ccffcc"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4207..4328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(4377..4393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4401..4417
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(4425..4455)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 4425 .. 4431 / #ffffff / -10
                        2: 4432 .. 4449 / #ffffff
                        3: 4450 .. 4455 / #ffffff / -35"
     protein_bind    4470..4491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4779..5367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5538..6398)
                     /label=AmpR
                     /note="color: #993366; direction: LEFT"
     promoter        complement(6399..6503)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcatccctat cagtgataga gatcagatct
      841 ccctatcagt gatagagagc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccggactct agcgtttaaa cttaagcttg gtacccgggg atccaccatg ggttctagac
      961 tggacaagag caaagtcata aactctgctc tggaattact caatgaagtc ggtatcgaag
     1021 gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc
     1081 acgtgaagaa caagcgggcc ctgctcgatg ccctggcaat cgagatgctg gacaggcatc
     1141 atacccactt ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca
     1201 agtcattccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc
     1261 gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag
     1321 gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct
     1381 gcgtattgga ggatcaggag catcaagtag caaaagagga aagagagaca cctaccaccg
     1441 attctatgcc cccacttctg agacaagcaa ttgagctgtt cgaccatcag ggagccgaac
     1501 ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg
     1561 aaagcggcgg gccaaaaaag aagagaaagg ctagcggcgc cggcgccttt ttggctacta
     1621 cacttgaacg tattgagaag aactttgtca ttactgaccc aaggttgcca gataatccca
     1681 ttatattcgc gtccgatagt ttcttgcagt tgacagaata tagccgtgaa gaaattttgg
     1741 gaagaaactg caggtttcta caaggtcctg aaactgatcg cgcgacagtg agaaaaatta
     1801 gagatgccat agataaccaa acagaggtca ctgttcagct gattaattat acaaagagtg
     1861 gtaaaaagtt ctggaacctc tttcacttgc agcctatgcg agatcagaag ggagatgtcc
     1921 agtactttat tggggttcag ttggatggaa ctgagcatgt ccgagatgct gccgagagag
     1981 agggcgtgat gctggccaag aagaccgccg agaacatcga cgaggccgcc cgccgccgcg
     2041 gctaaaccgg tgcggccgca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2101 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2161 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2221 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2281 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2341 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2401 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcct cgagtctaga
     2461 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2521 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2581 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2641 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2701 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2761 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2821 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2881 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2941 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtacctag
     3001 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt ggccaaaaag
     3061 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     3121 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     3181 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     3241 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     3301 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     3361 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     3421 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     3481 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     3541 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     3601 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     3661 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     3721 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     3781 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     3841 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     3901 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     3961 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     4021 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
     4081 gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4141 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4201 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4261 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4321 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4381 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4441 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4561 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4621 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4681 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4741 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4801 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4861 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4921 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4981 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5041 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5101 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5161 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5221 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5281 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5341 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5401 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5461 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5521 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5581 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5641 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5701 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5761 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5821 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5881 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5941 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6001 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6061 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6121 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6181 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6241 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6301 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6361 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6421 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6481 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
