LOCUS       Exported                6044 bp ds-DNA     circular SYN 09-MAY-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pFRT/TO/DESTFLAGHA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  AUTHORS   Markus Landthaler
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 2018 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6044
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pFRT/TO/DESTFLAGHA"
     promoter        232..958
                     /label=CMV
                     /note="/dnas_title=CMV"
                     /note="/vntifkey=29"
     misc_feature    793..1881
                     /label=sequenced verified
                     /note="sequenced verified"
     protein_bind    818..859
                     /note="2x TetO2"
     CDS             993..1034
                     /codon_start=1
                     /label=V5 tag
                     /note="V5 tag"
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1032..1056
                     /label=attB1
                     /note="attB1"
     CDS             1059..1847
                     /codon_start=1
                     /label=APEX2-NES
                     /note="APEX2-NES"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADALQLPPLERLTLD*"
     misc_feature    complement(1849..1873)
                     /label=attB2
                     /note="attB2"
     primer_bind     complement(1908..1932)
                     /label=Primer 1
     primer_bind     1933..1956
                     /label=Primer 2
     polyA_signal    2018..2242
                     /label=BGH
                     /note="/dnas_title=BGH"
                     /note="/vntifkey=25"
     misc_recomb     2526..2573
                     /label=FRT
                     /note="/dnas_title=FRT"
                     /note="/vntifkey=86"
     CDS             2581..3601
                     /codon_start=1
                     /label=Hygromycin
                     /note="/dnas_title=Hygromycin"
                     /note="/vntifkey=4"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    3733..3863
                     /label=SV40 early
                     /note="/dnas_title=SV40 early"
                     /note="/vntifkey=25"
     rep_origin      complement(4246..4919)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/dnas_title=pUC ori"
                     /note="/vntifkey=33"
     misc_feature    complement(5064..5924)
                     /label=Ampicillin
                     /note="/dnas_title=Ampicillin"
                     /note="/vntifkey=21"
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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tgggtaagcc tatccctaac cctctcctcg
     1021 gtctcgattc aacaagtttg tacaaaaaag caggcttcgg aaagtcttac ccaactgtga
     1081 gtgctgatta ccaggacgcc gttgagaagg cgaagaagaa gctcagaggc ttcatcgctg
     1141 agaagagatg cgctcctcta atgctccgtt tggcattcca ctctgctgga acctttgaca
     1201 agggcacgaa gaccggtgga cccttcggaa ccatcaagca ccctgccgaa ctggctcaca
     1261 gcgctaacaa cggtcttgac atcgctgtta ggcttttgga gccactcaag gcggagttcc
     1321 ctattttgag ctacgccgat ttctaccagt tggctggcgt tgttgccgtt gaggtcacgg
     1381 gtggacctaa ggttccattc caccctggaa gagaggacaa gcctgagcca ccaccagagg
     1441 gtcgcttgcc cgatcccact aagggttctg accatttgag agatgtgttt ggcaaagcta
     1501 tggggcttac tgaccaagat atcgttgctc tatctggggg tcacactatt ggagctgcac
     1561 acaaggagcg ttctggattt gagggtccct ggacctctaa tcctcttatt ttcgacaact
     1621 catacttcac ggagttgttg agtggtgaga aggaaggtct ccttcagcta ccttctgaca
     1681 aggctctttt gtctgaccct gtattccgcc ctctcgttga caaatatgca gcggacgaag
     1741 atgccttctt tgctgattac gctgaggctc accaaaagct ttccgagctt gggtttgctg
     1801 atgccctgca gctgcctccc ctggagcgcc tgaccctgga ctaatagaac ccagctttct
     1861 tgtacaaagt ggttgatatc cagcacagtg gcggccgctc aagtctagag ggcccgcggt
     1921 tcgaaggtaa ccggtaccgg atccgcggcc gcatagataa ctgatccagt gtgcctcgag
     1981 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2041 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2101 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2161 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2221 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2281 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2341 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2401 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2461 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2521 acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc
     2581 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     2641 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     2701 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     2761 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     2821 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     2881 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg
     2941 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     3001 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     3061 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     3121 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     3181 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     3241 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     3301 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg
     3361 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     3421 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     3481 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     3541 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     3601 tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3661 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3721 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3781 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3841 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     3901 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4441 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4741 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4981 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5041 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5101 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5161 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5221 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5281 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5341 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5401 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5461 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5521 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5581 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5641 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5701 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6001 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaa
//
