LOCUS       Exported                6450 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 6450)
  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..6450
                     /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..1910
                     /label=sequenceVerified Clone1 selected
                     /note="sequenceVerified Clone1 selected"
     protein_bind    818..859
                     /note="2x TetO2"
     primer_bind     complement(963..1001)
                     /label=Primer 4
     CDS             990..993
                     /codon_start=1
                     /label=FLAGHA
                     /note="/dnas_title=FLAGHA"
                     /note="/vntifkey=4"
                     /translation="M"
     CDS             993..1034
                     /codon_start=1
                     /label=V5
                     /note="/dnas_title=V5"
                     /note="/vntifkey=4"
                     /translation="GKPIPNPLLGLDST"
     CDS             1029..1031
                     /codon_start=1
                     /label=FLAGHA originB
                     /note="FLAGHA originB"
                     /translation="S"
     misc_feature    1032..1056
                     /label=AttB1
                     /note="AttB1"
     gene            1059..1433
                     /label=H2B
                     /note="H2B"
     misc_feature    complement(1320..2376)
                     /label=sequenceVerifiedRev
                     /note="sequenceVerifiedRev"
     misc_feature    1440..1484
                     /label=HaloTag 7 C-terminus linker
                     /note="HaloTag 7 C-terminus linker"
     primer_bind     complement(1455..1499)
                     /label=Primer 5
     misc_feature    1455..1475
                     /label=TEV protease recognition sequence
                     /note="TEV protease recognition sequence"
     primer_bind     complement(1461..1498)
                     /label=Primer 5
     CDS             1485..2237
                     /codon_start=1
                     /label=APEX2
                     /note="APEX2"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA*"
     misc_feature    complement(2239..2263)
                     /label=AttB2
                     /note="AttB2"
     primer_bind     complement(2298..2322)
                     /label=Primer 1
     primer_bind     2323..2346
                     /label=Primer 2
     polyA_signal    2408..2632
                     /label=BGH
                     /note="/dnas_title=BGH"
                     /note="/vntifkey=25"
     misc_recomb     2916..2963
                     /label=FRT
                     /note="/dnas_title=FRT"
                     /note="/vntifkey=86"
     CDS             2971..3991
                     /codon_start=1
                     /label=Hygromycin
                     /note="/dnas_title=Hygromycin"
                     /note="/vntifkey=4"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4123..4253
                     /label=SV40 early
                     /note="/dnas_title=SV40 early"
                     /note="/vntifkey=25"
     rep_origin      complement(4636..5309)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/dnas_title=pUC ori"
                     /note="/vntifkey=33"
     misc_feature    complement(5454..6314)
                     /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 caggcttccc agagccagcg aagtctgctc
     1081 ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga
     1141 agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg
     1201 tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg
     1261 acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga
     1321 ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca
     1381 agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct aaggtttctc
     1441 tcgagccaac cactgaggat ctgtactttc agagcgataa cgatggaaag tcttacccaa
     1501 ctgtgagtgc tgattaccag gacgccgttg agaaggcgaa gaagaagctc agaggcttca
     1561 tcgctgagaa gagatgcgct cctctaatgc tccgtttggc attccactct gctggaacct
     1621 ttgacaaggg cacgaagacc ggtggaccct tcggaaccat caagcaccct gccgaactgg
     1681 ctcacagcgc taacaacggt cttgacatcg ctgttaggct tttggagcca ctcaaggcgg
     1741 agttccctat tttgagctac gccgatttct accagttggc tggcgttgtt gccgttgagg
     1801 tcacgggtgg acctaaggtt ccattccacc ctggaagaga ggacaagcct gagccaccac
     1861 cagagggtcg cttgcccgat cccactaagg gttctgacca tttgagagat gtgtttggca
     1921 aagctatggg gcttactgac caagatatcg ttgctctatc tgggggtcac actattggag
     1981 ctgcacacaa ggagcgttct ggatttgagg gtccctggac ctctaatcct cttattttcg
     2041 acaactcata cttcacggag ttgttgagtg gtgagaagga aggtctcctt cagctacctt
     2101 ctgacaaggc tcttttgtct gaccctgtat tccgccctct cgttgacaaa tatgcagcgg
     2161 acgaagatgc cttctttgct gattacgctg aggctcacca aaagctttcc gagcttgggt
     2221 ttgctgatgc ctaatagaac ccagctttct tgtacaaagt ggttgatatc cagcacagtg
     2281 gcggccgctc aagtctagag ggcccgcggt tcgaaggtaa ccggtaccgg atccgcggcc
     2341 gcatagataa ctgatccagt gtgcctcgag tctagagggc ccgtttaaac ccgctgatca
     2401 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2461 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2521 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2581 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2641 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2701 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2761 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2821 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2881 aaaaaacttg attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat
     2941 tctctagaaa gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc
     3001 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     3061 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     3121 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     3181 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     3241 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     3301 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     3361 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     3421 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     3481 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     3541 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     3601 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     3661 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     3721 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     3781 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     3841 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     3901 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     3961 cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc
     4021 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4081 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4261 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4321 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4381 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4501 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4561 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4861 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5101 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5401 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5461 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5521 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5581 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5641 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5701 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5761 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5821 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5881 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5941 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6001 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6061 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6301 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6421 acatttcccc gaaaagtgcc acctgacgtc
//
