LOCUS       pLC_ZsGreen_P2A_        7614 bp    DNA     circular UNA 13-MAR-2018
DEFINITION  pLC_P2A_Blast-based expression vector for ZsGreen.
ACCESSION   urn.local...1527182838683.49
VERSION     urn.local...1527182838683.49
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    6235..6242
                     /note="Geneious type: Editing History Replacement"
                     /label="GATCGATCC"
     gene            2623..3483
                     /label="Gene_1"
     misc_recomb     416..449
                     /label="LoxP"
     misc_feature    3616^3617
                     /note="Geneious type: origin"
                     /label="origin"
     misc_feature    3828..4432
                     /note="Geneious type: promoter prokaryotic"
                     /label="CMV promoter/enhancer"
     misc_feature    6243..6833
                     /note="Geneious type: promoter uukaryotic"
                     /label="CMV"
     rep_origin      1805..2478
                     /label="pUC"
     source          order(<6859..>7548,<6859..>7548)
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /label="source recombinant plasmid"
     misc_feature    6205..6234
                     /note="Geneious type: GibsonFlank"
                     /label="Gibson_5'overhang_KillBam"
     misc_feature    6826..6855
                     /note="Geneious type: GibsonFlank"
                     /label="5'Gibson"
     LTR             1113..1802
                     /label="3' SIN-LTR"
     LTR             4451..5125
                     /label="5' LTR"
     CDS             <6856..>7548
                     /codon_start=1
                     /note="ZsGreen"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI
                     NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG
                     AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI
                     LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH
                     LTEHAIASGSALP"
                     /label="ZsGreen"
     CDS             1..396
                     /label="BlastR"
     CDS             7549..7614
                     /label="P2A"
     primer_bind     7568..18
                     /label="3910Zs_BlastF"
     primer_bind     6205..6262
                     /label="3883CMVF_Pac"
     primer_bind     6826..6875
                     /label="3888_CMV_ZsGreenF"
     misc_feature    complement(7529..7597)
                     /note="Geneious type: primer_bind_reverse"
                     /label="3887_ZsGreen_P2AR"
     misc_feature    complement(6836..6885)
                     /note="Geneious type: primer_bind_reverse"
                     /label="3885CMVR_ZsG"
     misc_feature    complement(377..426)
                     /note="Geneious type: primer_bind_reverse"
                     /label="3911Blast_pL_R"
     misc_feature    complement(465..483)
                     /note="Geneious type: primer_bind_reverse"
                     /label=258
     misc_feature    2623..3483
                     /label="AmpR"
     misc_feature    5155..6012
                     /label="Psi"
     misc_feature    504..1093
                     /label="WRE"
     misc_feature    6038..6215
                     /label="FLAP"
     misc_feature    1622..1719
                     /label="R"
     misc_feature    4451..4548
                     /label="R"
     misc_feature    7481..7500
                     /label="AJ's Seq1"
     misc_feature    397..402
                     /label="Kept EcoRI site in - need to restore with Gibson"
     misc_feature    7363..7368
                     /label="K6-5p 2-8A"
     misc_feature    6850..6855
                     /label="Kozak Sequence"
     misc_feature    6234
                     /label="cut next 4 nts off - overhang after BamHi
                     digestions"
     misc_feature    6235..6242
                     /note="Geneious type: restriction site"
                     /label="PacI"
     misc_feature    397..402
                     /note="Geneious type: restriction site"
                     /label="EcoRI"
     misc_feature    7549..7554
                     /note="Geneious type: restriction site"
                     /label="BamHI"
     misc_feature    6834..6839
                     /note="Geneious type: restriction site"
                     /label="NheI"
     misc_feature    3847..3852
                     /note="Geneious type: restriction site"
                     /label="MluI"
     misc_feature    6843..6848
                     /note="Geneious type: restriction site"
                     /label="AgeI"
ORIGIN      
        1 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
       61 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
      121 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
      181 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
      241 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
      301 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
      361 gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgtcgaggg acctaataac
      421 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
      481 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
      541 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
      601 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
      661 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
      721 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
      781 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
      841 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
      901 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
      961 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     1021 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     1081 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     1141 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     1201 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     1261 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     1321 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     1381 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     1441 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     1501 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     1561 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     1621 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     1681 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     1741 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     1801 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     3661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     3721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     3781 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     3841 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     3901 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3961 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     4021 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     4081 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     4141 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     4201 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     4261 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4321 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     4381 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
     4441 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4501 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4561 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4621 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     4681 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     4741 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     4801 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     4861 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     4921 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     4981 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5041 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5101 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg cgctgatctt
     5161 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     5221 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     5281 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     5341 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     5401 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     5461 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     5521 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     5581 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     5641 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     5701 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     5761 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     5821 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     5881 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     5941 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     6001 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca
     6061 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     6121 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     6181 gggtttatta cagggacagc agagatccag tttggttagt accgggcccg agagttaatt
     6241 aattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     6421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     6481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     6781 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     6841 ctaccggtcg ccaccatggc ccagtccaag cacggcctga ccaaggagat gaccatgaag
     6901 taccgcatgg agggctgcgt ggacggccac aagttcgtga tcaccggcga gggcatcggc
     6961 taccccttca agggcaagca ggccatcaac ctgtgcgtgg tggagggcgg ccccttgccc
     7021 ttcgccgagg acatcttgtc cgccgccttc atgtacggca accgcgtgtt caccgagtac
     7081 ccccaggaca tcgtcgacta cttcaagaac tcctgccccg ccggctacac ctgggaccgc
     7141 tccttcctgt tcgaggacgg cgccgtgtgc atctgcaacg ccgacatcac cgtgagcgtg
     7201 gaggagaact gcatgtacca cgagtccaag ttctacggcg tgaacttccc cgccgacggc
     7261 cccgtgatga agaagatgac cgacaactgg gagccctcct gcgagaagat catccccgtg
     7321 cccaagcagg gcatcttgaa gggcgacgtg agcatgtacc tgctgctgaa ggacggtggc
     7381 cgcttgcgct gccagttcga caccgtgtac aaggccaagt ccgtgccccg caagatgccc
     7441 gactggcact tcatccagca caagctgacc cgcgaggacc gcagcgacgc caagaaccag
     7501 aagtggcacc tgaccgagca cgccatcgcc tccggctccg ccttgcccgg atccggcgca
     7561 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accg
//
