LOCUS       pSAP5::Tag-RFPt-_3UTR_Meganuclease        9249 bp    DNA     linear   UNA 19-JAN-2019
DEFINITION  LR.
ACCESSION   urn.local...54b-dt34muk
VERSION     urn.local...54b-dt34muk
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     primer_bind     205..221
                     /standard_name="M13 Reverse"
     primer_bind     239..256
                     /standard_name="SP6 Primer"
     misc_feature    339..356
                     /note="Geneious type: motif"
                     /standard_name="Meganuclease Site"
     misc_feature    362..364
                     /note="Geneious type: Editing History Replacement"
                     /standard_name="TAA"
     regulatory      365..3473
                     /note="Geneious type: promoter"
                     /standard_name="pSAP5 3kb"
     primer_bind     365..392
                     /standard_name="pSap5 For"
     misc_feature    complement(610..632)
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 21"
     misc_feature    612..634
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 5"
     misc_feature    complement(1059..1081)
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 20"
     misc_feature    1124..1146
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 6"
     misc_feature    1150..1172
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 7"
     misc_feature    complement(1680..1702)
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 19"
     misc_feature    complement(1985..2007)
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 18"
     misc_feature    2530..2552
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 8"
     misc_feature    complement(3447..3473)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="pSap5 Rev"
     misc_feature    3447..3469
                     /note="Geneious type: CRISPR"
                     /standard_name="CRISPR guide 9"
     CDS             3482..4195
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQT
                     MRIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYED
                     GGVLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEG
                     RTDMALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQH
                     EVAVARYCDLPSKLGHKLN"
                     /standard_name="TagRFP"
     misc_feature    4318..4439
                     /note="Geneious type: polyA_signal"
                     /note="SV40 polyadenylation signal"
                     /standard_name="SV40 poly(A) signal"
     primer_bind     complement(4355..4374)
                     /note="SV40 polyA, reverse primer"
                     /standard_name="SV40pA-R"
     primer_bind     4409..4428
                     /note="SV40 polyA terminator, reverse primer"
                     /standard_name="EBV-rev"
     rep_origin      complement(4446..>4448)
     exon            <4451..4453
                     /standard_name="Exon2"
     misc_feature    4451..4477
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="Sap5 3UTR FOR"
     3'UTR           4451..5584
                     /standard_name="SAP5 3UTR 1kb"
     misc_feature    complement(5557..5584)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="Sap5 3UTR Rev "
     misc_feature    complement(5593..5610)
                     /note="Geneious type: motif"
                     /standard_name="Meganuclease site"
     primer_bind     complement(5682..5701)
                     /standard_name="T7 Primer"
     primer_bind     complement(5708..5724)
                     /standard_name="M13 Forward -20"
     primer_bind     complement(5728..5744)
                     /standard_name="M13 Forward -40"
     gene            6637..7431
                     /standard_name="Ampicillin R"
     gene            7449..8309
                     /standard_name="Kanamycin R"
ORIGIN      
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttgata gggataacag ggtaatttaa
      361 tcgcgtaaca acaccaagcc atgtgttagc ccaataattc aaaatgaatg aatttacttc
      421 aattgcagca ggttatgcag tatcccttat ttcagcaaat gagcagaaaa tcttatttga
      481 ttttcaaaaa gcctttgatt atgttattca taacattgtt cttatgaacc taaaataaaa
      541 gatgaaacag tatagttgtt agcttctttt taaataacag aacacaaagg ataaaataag
      601 ggatgggatc cgggtagtga agtgaatgtg ccggataccc gataatttac cggataccgg
      661 ctaatatatt gttcaactat tactagcgga aacagaaaaa tgactttaaa aacaatatat
      721 aaccttattt ttatgataaa taatgaggga atttagttaa atatgcctaa aatatgacca
      781 tccatttttc aaaataatca attttagcaa tatataacgt ttttcaatta cgagaacaag
      841 ataaaaacca aacaaaaata attaaataaa aaagtcaatt ttttaaagct actaaattat
      901 aataaaagta cataactgtc catacattat tgtggttaag atttctgtca taggctagga
      961 gttttcttag catagaaaaa tatctaccaa ctgcagccga tgttgattga cattttcgta
     1021 gcttggcgta caatttagat taactgtcca tccccttgcc atccattata gcagtaaagc
     1081 cggtattttc tattcgtgac ttgaaatatg caattatttc acaggcatca tttccaaaaa
     1141 tcaagggtgg gagtaataac aacatctggt ggaatttcta atctgtcaaa catttaacaa
     1201 ctttgatttt ttatgatagc agcattaatt gaagcaatta gattgtgtag cctcatatgt
     1261 ttttgccgga atacgcaagg tacaattctg caacacgtgt gccttgtaga atccataaaa
     1321 taatcatgta tattgttctt tattattttt tccactgcca gctcttatgg ccagtcctag
     1381 aaacgcgttg atcagtgagt cgcgtcgagt taataattag ctaggaatta aatttctcgg
     1441 gggttagaca aggaagtata ttaattgaaa ttgaatgaat ataaatgtct tagttcaaat
     1501 gatataaagc agtcacaatc caacattaat ttttaatttt ttttcttaat tttaatgaaa
     1561 ctatgctaac atatcacttt tatgacattt cagccacaaa taccgtattt ttcaaaaaac
     1621 aaaaatatga ctttttatga ctttatactt cctatttaat tgcaggccag atctgaaaac
     1681 catcaaattt taagcagact ccaaaacgga ccaacagaaa aaaatgttac ttttatgact
     1741 cttagttccc ctaattttta cagaagaaca acctcatttc tcaattaata tcgacctctt
     1801 gcgaatgaat ataaataatt attaaataga ctatgtcctg atgtatcaga aaaatttgtt
     1861 ttgtcgcatg acaacttcca tatcatggga aaatataaat tcctttttaa ttcaagtttt
     1921 atctcagaat atattatccg gtaaaatccg gtaaacaagt acaattttac cggataccgg
     1981 atatccagat caagttgtgt aattgccgat tttaccagat accggattct ggcacatccc
     2041 tagataaaac accttgtgga attcatttag aactgaacat aagagactaa attaggacca
     2101 tatttgtctg ttttattcat aattatgttg ctagggaatt ggtaccaaag aaataaattt
     2161 gtcgatgaca cgacagcctg agattctatt aatgaagaac aaagtcacat aattattgat
     2221 atgataaagg actggtgtac tgaaaatgat gttgtaatta atactaacaa agctagagaa
     2281 attacaaatt taccacaaga aatagaaata gattatagaa taataaaaag aagcaagaca
     2341 aaaaaattta ttgtagtaca gttcaagaca gtctgaaatg ggatgaacat ataaatcaac
     2401 taattggtaa agcaactaag ataaagtcat ttcttcaaac aatttatctt cagtggtata
     2461 ccggtaaaag atctttacat atttacttcg ctatgatcag atcatcatta gaatactgtt
     2521 gtcaataacg gcatagcacg ataacagaaa ggcaaaagtt taaagactga aagcatacaa
     2581 aggagagttt ttagaattat aaaacctaac ttacattact ttgaagcaca tgaatttatt
     2641 aaatttttaa atgtaaatgt ttatttgaat attaatgaca aaaattatgg ttaaatataa
     2701 aaaatcccca ccgtccatta agctatctaa gacaaaataa taacgaatgc agtcatgatt
     2761 atccaattct aactcttagg gaaaggcttc aaataggttg agtgattcat ttatactttc
     2821 aaagaaaagg aaagatttta cggcgtccta catttaattt gtttttgttt cttttttatg
     2881 ttctcttgat attattcaag tctaaagttt gcaataaaaa ctaatatatt tactattact
     2941 attactttaa aaggcttcaa aatccaattg ttcttcctac atgatatttt gcttgcaaaa
     3001 gaaagaatca ttgtagctaa aatgtcgttg aaaagtgttc actaataaat tccaagtacg
     3061 tgctcgaatt tgaaaacatt tgctaaagca aaaaatatat aaagtcttcg cttccaaatc
     3121 attagctttt gtcaaatttg gatatttttt gaaaatttgt ttcatgttat tcacaaaatt
     3181 gccactttaa gacattctta tcaataaaaa attttctgat ttctaaatgg acgtgtacat
     3241 gaatacagtt acttgcaaac atagcattga taaactaaca aattagtcat tctaaattta
     3301 tggaaaggtg tgcacattca ataagctatt agttccgtaa aaatttatat acataatgat
     3361 taagtaacaa aaagcaaaca caatcaaaat ttcaattatt taagtaatgt aacttattga
     3421 caaacatcta tgtataaata gagcatggat aaatagcatc agtattagga aagggcgcgc
     3481 catggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga
     3541 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     3601 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     3661 catcctggct accagcttca tgtacggcag cagaaccttc atcaaccaca cccagggcat
     3721 ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     3781 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3841 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     3901 aacactcggc tgggaggcca acaccgagat gctgtacccc gctgacggcg gcctggaagg
     3961 cagaaccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttcaagac
     4021 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     4081 ccacagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     4141 ggctgtggcc agatactgcg acctccctag caaactgggg cacaaactta attaaagcgg
     4201 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4261 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     4441 ggcgtaaagg taatcagaaa ttaagaatca atagaagtaa acgtagtagt tttctaattg
     4501 aaatatacat atctaatttt gcttttcaac ctgatcaaat tggattctga agttttgcaa
     4561 ttactattgt agatatattt attaaatatc attttgccat tttttcattg ggatgaataa
     4621 acttccgatc tcattaaaag tcttgactaa agtctctgac tgtattttta acttttcaat
     4681 tttgatttta gttaatttaa aaacaataca aaagcaatat tataagaatt acactactaa
     4741 gcatttacta ataatgtatt ttatcaatac attgaactcc atgtctctat gtttacattt
     4801 agagacttga aatgggtgaa aatagaattc atagaaaaat ctaaaatatg ttgttcacag
     4861 tagtctacat attcatacaa tgtgaaataa tttctaaaga gcatttacct tatatctcat
     4921 attcaatgac caagtttcat cgtaaatagt tattgttgaa ggtgtctaat tgaaattcag
     4981 gcaaagatat gctatatatt cgtaataata taaattcttt gtttggatgt atatgctaag
     5041 ttaatcacaa acattcatgt taatacaaat aatttttatg cacggaatca caatgcaaag
     5101 atgtatattt ttcggatacc gaatgaagac catattttca tttagaaaaa agttaacctt
     5161 tgcctgcttt tatcaaatta caccatatag taatgagtat gttaaatgtc tattcaaagt
     5221 aaacaacaaa caaattacgt gaatcagcta taaaaaatat taaatattct catcgtcgtt
     5281 attaagaatg aagatagact tgcgagattt tattgtcaga aatatatgct atttctattt
     5341 ttttatgaaa ggtgacattt caattaataa ggtcaacagt aaaaaaagcc ttaaacaaat
     5401 ttcatttaca caaatatagc agtaaaaaat ccaaaagcta ataaagcgcg tggaaaaaaa
     5461 attttaaaag aacgaataaa atgaccataa tctaccaaat atttagtagt ttttaaggat
     5521 gcaattatct attcttttca tatatttatg aagcatgcat ttatattcta gtattgtgat
     5581 atacggccgg ccattaccct gttatcccta tcaagggcga attctgcaga tatccatcac
     5641 actggcggcc gctcgagcat gcatctagag ggcccaattc gccctatagt gagtcgtatt
     5701 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5761 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5821 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     5881 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5941 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6001 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6061 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6121 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6181 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6241 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6301 aattcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa
     6361 cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc
     6421 gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt
     6481 ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag
     6541 ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca
     6601 agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6661 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6721 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6781 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6841 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6901 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc ccaccttgct
     6961 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     7021 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7081 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7141 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7201 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7261 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7321 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7381 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattgaaaaa
     7441 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7501 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7561 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7621 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7681 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7741 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7801 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7861 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7921 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7981 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     8041 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     8101 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     8161 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     8221 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8281 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8341 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8401 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8461 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8581 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     9001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     9061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     9121 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     9181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     9241 aagcggaag
//
