LOCUS       fluoPEER-GFP>BFP        5986 bp ds-DNA     circular     20-APR-2022
DEFINITION  .
SOURCE      synthetic DNA construct                                            
  ORGANISM  synthetic DNA construct                                            
FEATURES             Location/Qualifiers
     rep_origin      110..698
                     /label="ori"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     599..618
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     enhancer        873..1176
                     /label="CMV enhancer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     promoter        1177..1380
                     /label="CMV promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     1331..1351
                     /label="CMV-F"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          complement(1408..1427)
                     /label="pJet-mCherry-Rv"
                     /note="sequence: catggtggcgaccggtagcg"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     regulatory      1419..1428
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             1425..2141
                     /label="EGFP"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             1425..2141
                     /label="Translation 1425-2141"
     primer_bind     complement(1470..1491)
                     /label="EGFP-N"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     complement(1731..1750)
                     /label="EXFP-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             2142..2228
                     /label="Translation 2142-2228"
     CDS             2157..2213
                     /label="P2A"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     CDS             2220..2285
                     /label="3xFLAG"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             2229..2615
                     /label="Translation 2229-2615"
     misc_feature    2565..2586
                     /label="Fw-for-mKate"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             2616..2745
                     /label="Translation 2616-2745"
     misc_feature    2622..2667
                     /label="GFP edit region"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    complement(2639..2658)
                     /label="pegRNA spacer binding site"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             2683..2739
                     /label="P2A"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(2725..2745)
                     /label="Rv for mKate"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          2746..2765
                     /label="mCherry Fw"
                     /note="sequence: gtgagcaagggcgaggagga"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             2746..3453
                     /label="mCherry"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             2746..3453
                     /label="Translation 2746-3453"
     primer_bind     complement(2888..2906)
                     /label="mCherry-R"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer_bind     3156..3175
                     /label="mCherry-F"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     polyA_signal    3586..3707
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     complement(3623..3642)
                     /label="SV40pA-R"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     3677..3696
                     /label="EBV-rev"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     rep_origin      complement(3714..4169)
                     /label="f1 ori"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     complement(3851..3872)
                     /label="F1ori-F"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer_bind     4063..4082
                     /label="F1ori-R"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        4196..4300
                     /label="AmpR promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        4302..4659
                     /label="SV40 promoter"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     complement(4325..4345)
                     /label="pBABE 3'"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     rep_origin      4510..4645
                     /label="SV40 ori"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     4572..4591
                     /label="SV40pro-F"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     CDS             4694..5488
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     complement(4748..4767)
                     /label="Neo-R"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     5358..5377
                     /label="Neo-F"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer_bind     complement(5676..5695)
                     /label="TK-pA-R"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     polyA_signal    5720..5767
                     /label="HSV TK poly(A) signal"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
ORIGIN
        1 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
       61 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
      121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      241 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      361 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
      421 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
      481 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
      541 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      661 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      721 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      781 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
      841 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      901 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      961 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1021 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1081 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1141 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1201 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1261 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1321 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1381 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
     1441 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1501 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1561 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1621 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1681 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1741 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1801 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1861 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1921 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1981 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2041 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtccttaag gagttcgtga
     2101 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggaggc agcggcgcca
     2161 ccaacttttc cctgctcaag caggccggcg acgtggaaga gaatcccggc cccggtactg
     2221 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg
     2281 acaaggccgg atctcctggt ccgaccccca gtggcactaa cgtgggatcc tcactctctg
     2341 acgaggactt caaggctgtt tttggcatga cccgctctgc ctttgccaac ttgcccttgt
     2401 ggaaacagca gaacctcaag aaggagaaag gactcttcgg atcagggcgc tctcccagca
     2461 aagcagtggc cgcccgggcg gcgggatcca ctgtccggca gaggaaaaat gccagctgtg
     2521 ggacaaggag tgcaggccgc acaacctcgg caggcaccgg ggggatgtgg cgattctaca
     2581 cagaagattc acctgggctc aaagttggcc ctaggtcgac cgtgaccacc ctgaacctac
     2641 ggcgtgcagt gcttcagccg ctaccccggt accggaagcg gagctactaa cttcagcctg
     2701 ctgaagcagg ctggagacgt ggaggagaac cctggacctc tcgaggtgag caagggcgag
     2761 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2821 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2881 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2941 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     3001 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     3061 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     3121 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     3181 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     3241 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     3301 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     3361 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     3421 tccaccggcg gcatggacga gctgtacaag taaactggat ccaccggatc tagataactg
     3481 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3541 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3601 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3721 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3781 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3841 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3901 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3961 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4021 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4081 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4141 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4261 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4321 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4381 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4501 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4561 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4621 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4681 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4741 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4801 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4861 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4921 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4981 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5041 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5101 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5161 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5221 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5281 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5341 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5401 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5461 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5521 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5581 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5641 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5701 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5761 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5821 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5881 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5941 tactcatata tactttagat tgatttaaaa cttcattttt aattta
//