LOCUS       pJet-CMV-GFP-P2A-15xILE 6177 bp ds-DNA     circular     26-FEB-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     primer_bind     304..324
                     /label="T7"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    310..332
                     /label="pJET Fw primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    348..353
                     /label="XhoI site"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    371..3575
                     /label="Insert"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          372..407
                     /label="pJet-stall-Fw-L"
                     /note="sequence: attggggggtacagtgcaggggaaagaatagtagac"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          380..402
                     /label="pJet-stall-Fw"
                     /note="sequence: gtacagtgcaggggaaagaatag"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    493..797
                     /label="CMV Enhancer"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     promoter        798..1001
                     /label="CMV promoter"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          952..972
                     /label="CMV-Fw"
                     /note="sequence: cgcaaatgggcggtaggcgtg"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          1132..1171
                     /label="Stal-Rep-for-64-infus-Fw-l"
                     /note="sequence: gaattctgcagtcgacAtggtgagcaagggcgaggagctg"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    1148..1148
                     /label="SapI overlap"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    1148..1148
                     /label="MCS"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     CDS             complement(1148..1148)
                     /label="Eco47I/T7"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             1148..1150
                     /label="Translation 1148-1150"
     misc_feature    1149..1151
                     /label="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    1149..1152
                     /label="overlap"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             1149..1864
                     /label="EGFP"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             1151..3268
                     /label="Translation 1151-3268"
     primer_bind     complement(1193..1214)
                     /label="EGFP-N"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer_bind     complement(1454..1473)
                     /label="EXFP-R"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     CDS             1880..1936
                     /label="P2A"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     CDS             1943..2008
                     /label="3xFLAG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    2009..2356
                     /label="Spacer-5'"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(2337..2377)
                     /label="Vector pM-GFP_R1"
                     /note="sequence: GATCACCACCTGCAACGTCATggtcgccatggcggtcgacc"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    2358..2360
                     /label="S.E."
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    2363..2461
                     /label="50x Isoleucine (new GB)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    2471..2536
                     /label="P2A"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(2541..2561)
                     /label="GB_Ile/Leu_SapI_R"
                     /note="sequence: GCTATGGCTCTTCGcGTGCTTGGACTGGGCCATGA"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          2541..2584
                     /label="Vector pM-GFP_F1"
                     /note="sequence: GATCACCACCTGCGATCCACGtgagcaagggcgaggaggataac"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    2558..2560
                     /label="S.E."
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             2561..3268
                     /label="mCherry"
                     /ApEinfo_revcolor=#f6989d
                     /ApEinfo_fwdcolor=#f6989d
     primer_bind     complement(2703..2721)
                     /label="mCherry-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          complement(2763..2782)
                     /label="p1013-Seq7-R"
                     /note="sequence: CTTGGAGCCGTACATGAACT"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     2971..2990
                     /label="mCherry-F"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     polyA_signal    3401..3520
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     complement(3438..3457)
                     /label="SV40pA-R"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer_bind     3492..3511
                     /label="EBV-rev"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(3492..3538)
                     /label="Stal-Rep-for-64-infus-Rv-l"
                     /note="sequence: gctgtctagaCTCGAGtaagatacattgatgagtttggacaaaccac"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    3519..3520
                     /label="overlap SapI"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    3520..3520
                     /label="overlap"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(3540..3561)
                     /label="pJet-stall-Rv"
                     /note="sequence: cttaatgttaacgaccggtgag"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(3540..3574)
                     /label="pJet-stall-Rv-L"
                     /note="sequence: ggcactaggctagcttaatgttaacgaccggtgag"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    3580..3583
                     /label="XbaI site"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(3608..3631)
                     /label="pJET Rv seq"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        complement(4009..4039)
                     /label="lac UV5 promoter"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     rep_origin      complement(4348..5030)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(5128..5787)
                     /label="AmpR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
      421 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgataagc
      481 ttgggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      541 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      601 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      661 gtatcatatg ccaagtacgc cccctattga cggtcaatga cggtaaatgg cccgcctggc
      721 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      781 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      841 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      901 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      961 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aAccgtcaga
     1021 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagctag
     1081 aggatccccc gggctgcagg aattcggcgc gccacgcgta ccgatagctt cgaattctgc
     1141 agtcgacAtg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1201 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1261 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1321 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1381 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1441 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1501 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1561 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1621 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1681 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1741 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1801 catggtcctt aaggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1861 caagtccgga ggcagcggcg ccaccaactt ttccctgctc aagcaggccg gcgacgtgga
     1921 agagaatccc ggccccggta ctgactacaa agaccatgac ggtgattata aagatcatga
     1981 catcgattac aaggatgacg atgacaaggc cggatctcct ggtccgaccc ccagtggcac
     2041 taacgtggga tcctcactct ctgacgagga cttcaaggct gtttttggca tgacccgctc
     2101 tgcctttgcc aacttgccct tgtggaaaca gcagaacctc aagaaggaga aaggactctt
     2161 cggatcaggg cgctctccca gcaaagcagt ggccgcccgg gcggcgggat ccactgtccg
     2221 gcagaggaaa aatgccagct gtgggacaag gagtgcaggc cgcacaacct cggcaggcac
     2281 cggggggatg tggcgattct acacagaaga ttcacctggg ctcaaagttg gccctaggtc
     2341 gaccgccatg gcgaccATGA TTATtGGCAG TATCAGCATt ATCTCAGGTA TTGGCGGATC
     2401 TAGTGGGGGT ATtGGCGGGG GCGGCATAAT CGGTGGAGGC ATTATCATAT CTATTATCAT
     2461 aGGCACCGGT GGAAGCGGAG CTACTAACTT CAGCCTGCTG AAGCAGGCTG GAGACGTGGA
     2521 GGAGAACCCT GGACCTGAAT TCATGGCCCA GTCCAAGCAC gtgagcaagg gcgaggagga
     2581 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2641 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2701 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2761 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2821 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2881 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2941 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     3001 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     3061 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     3121 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     3181 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     3241 cggcggcatg gacgagctgt acaagtaaac tggatccacc ggatctagat aactgatcat
     3301 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3361 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3421 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3481 gcattctagt tgtggtttgt ccaaactcat caatgtatct taCTCGAGtc tagacagcgc
     3541 tcaccggtcg ttaacattaa gctagcctag tgccatcttt ctagaagatc tcctacaata
     3601 ttctcagctg ccatggaaaa tcgatgttct tcttttattc tctcaagatt ttcaggctgt
     3661 atattaaaac ttatattaag aactatgcta accacctcat caggaaccgt tgtaggtggc
     3721 gtgggttttc ttggcaatcg actctcatga aaactacgag ctaaatattc aatatgttcc
     3781 tcttgaccaa ctttattctg catttttttt gaacgaggtt tagagcaagc ttcaggaaac
     3841 tgagacagga attttattaa aaatttaaat tttgaagaaa gttcagggtt aatagcatcc
     3901 attttttgct ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt tgacatgttt
     3961 aaagtttaaa cctcctgtgt gaaattatta tccgctcata attccacaca ttatacgagc
     4021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4081 gttgcgctca ctgccaattg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4801 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6121 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcc
//