LOCUS       LifeAct-scramble        5461 bp ds-DNA     circular     26-FEB-2024
DEFINITION  .
KEYWORDS    "marker:AmpR, seq primer R, AmpR"
FEATURES             Location/Qualifiers
     polyA_signal    121..255
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     rep_origin      complement(494..1082)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             complement(1253..2044)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             complement(2045..2113)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        complement(2114..2218)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        2274..2588
                     /label="tight TRE promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    2282..2300
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2318..2336
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2353..2371
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2389..2407
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2425..2443
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2460..2478
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     protein_bind    2496..2514
                     /label="tet operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     primer          2562..2582
                     /label="EH-1"
                     /note="sequence: GCTCGTTTAGTGAACCGTCAG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          2594..2670
                     /label="EH-11 LifeAct Ultramer"
                     /note="sequence: AAACCGGTtatactGCGGCCGCCACCATGggcgtggcggatctgattaaGaaGttcgaaagcattagcaaagaagaa"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    2602..2607
                     /label="filler (loop)"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    2611..2622
                     /label="Kozak Sequence"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2623..2670
                     /label="LifeAct"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             2623..2670
                     /label="Translation 2623-2670"
                     /translation="GVADLIKKFESISKEE"
     primer          2643..2690
                     /label="EH-9"
                     /note="sequence: GaaGttcgaaagcattagcaaagaagaaggctctggttccggcACTAG"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     CDS             2671..2685
                     /label="linker"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     CDS             2692..3384
                     /label="scarlet-I"
                     /ApEinfo_revcolor="#ff2a75"
                     /ApEinfo_fwdcolor="#ff2a75"
     CDS             2692..3384
                     /label="Translation 2692-3384"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHSTGGMDELYK"
     primer          complement(2757..2794)
                     /label="EH-10"
                     /note="sequence: CCTCTCCTTCCCCTTCGATTTCGAAcTCGTGTCCGTTC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(2889..2908)
                     /label="EH-3"
                     /note="sequence: TAAATGCTCGGGAGCCATAC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    2931..2931
                     /label="Silent Mutation #1"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    3159..3159
                     /label="Silent Mutation #2"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    3321..3321
                     /label="Silent Mutation #3"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             3385..3402
                     /label="linker"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          3385..3444
                     /label="Primer 210"
                     /note="sequence: ggcagcggcagcggcagcatgattaccgcgccgcgcgaatttgattatctgaacctgcgc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    3403..3462
                     /label="MTED Scramble"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     CDS             3403..3462
                     /label="MTED scramble"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MITAPREFDYLNLRAGLSMT"
     primer          complement(3427..3486)
                     /label="Primer 211"
                     /note="sequence: CAACCCCGGAGCATGACGCGTtcaggtcatgctcaggcccgcgcgcaggttcagataatc"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     promoter        3480..3990
                     /label="hPGK promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(3580..3599)
                     /label="EH-4"
                     /note="sequence: GACGTGAAGAATGTGCGAGA"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             4000..4617
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             4618..4623
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4624..4659
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             4660..4662
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4663..4698
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             4699..4701
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             4702..4737
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             4738..4746
                     /label="rtTA-Advanced"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(4777..4794)
                     /label="seq primer R"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     polyA_site      4794..4849
                     /label="β-globin poly(A) signal"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     Primer          complement(4848..4867)
                     /label="Dent_Seq_11"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     complement(5208..5224)
                     /label="M13 rev"
                     /ApEinfo_revcolor="#a020f0"
                     /ApEinfo_fwdcolor="#a020f0"
     Primer          complement(5208..5224)
                     /label="Dent_Seq_14"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     protein_bind    5232..5248
                     /label="lac operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     promoter        complement(5256..5262)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(5263..5280)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     promoter        complement(5281..5286)
                     /label="lac promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     protein_bind    5301..5322
                     /label="CAP binding site"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     promoter        5381..115
                     /label="SV40 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     rep_origin      5427..101
                     /label="SV40 ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
ORIGIN
        1 TCCGCCCCAT GGCTGACTAA TTTTTTTTAT TTATGCAGAG GCCGAGGCCG CCTCGGCCTC
       61 TGAGCTATTC CAGAAGTAGT GAGGAGGCTT TTTTGGAGGC CTAGGCTTTT GCAAAAAGCT
      121 AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA ATAGCATCAC AAATTTCACA
      181 AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT CCAAACTCAT CAATGTATCT
      241 TATCATGTCT GGATCCGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC
      301 GTATTGGGCG CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC
      361 GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA
      421 ACGCAGGAAA GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG
      481 CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT
      541 CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA
      601 GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC
      661 TCCCTTCGGG AAGCGTGGCG CTTTCTCAAT GCTCACGCTG TAGGTATCTC AGTTCGGTGT
      721 AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG
      781 CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG
      841 CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT
      901 TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGGACAGT ATTTGGTATC TGCGCTCTGC
      961 TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG
     1021 CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC
     1081 AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT
     1141 AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA
     1201 AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT
     1261 GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT
     1321 GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG
     1381 CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG
     1441 CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA
     1501 ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG
     1561 CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG
     1621 GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT
     1681 CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA
     1741 TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG
     1801 GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC
     1861 CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG
     1921 GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA
     1981 TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG
     2041 GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT
     2101 GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC
     2161 TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA
     2221 CATTTCCCCG AAAAGTGCCA CCTGGTCGTC GACTGGCCGC GAATCGATAT GTCGAGTTTA
     2281 CTCCCTATCA GTGATAGAGA ACGTATGTCG AGTTTACTCC CTATCAGTGA TAGAGAACGA
     2341 TGTCGAGTTT ACTCCCTATC AGTGATAGAG AACGTATGTC GAGTTTACTC CCTATCAGTG
     2401 ATAGAGAACG TATGTCGAGT TTACTCCCTA TCAGTGATAG AGAACGTATG TCGAGTTTAT
     2461 CCCTATCAGT GATAGAGAAC GTATGTCGAG TTTACTCCCT ATCAGTGATA GAGAACGTAT
     2521 GTCGAGGTAG GCGTGTACGG TGGGAGGCCT ATATAAGCAG AGCTCGTTTA GTGAACCGTC
     2581 AGATCGCCTG GAGAAACCGG TtatactGCG GCCGCCACCA TGggcgtggc ggatctgatt
     2641 aaGaaGttcg aaagcattag caaagaagaa ggctctggtt ccggcACTAG TGTGAGTAAA
     2701 GGAGAGGCCG TTATAAAAGA GTTCATGCGG TTCAAGGTAC ACATGGAGGG ATCAATGAAC
     2761 GGACACGAgT TCGAAATCGA AGGGGAAGGA GAGGGCAGGC CCTATGAGGG TACtCAGACC
     2821 GCCAAACTCA AAGTGACAAA GGGAGGCCCT TTGCCTTTCT CTTGGGACAT CTTGTCACCA
     2881 CAATTTATGT ATGGCTCCCG AGCATTTATA AAACACCCAG CAGACATACC tGATTACTAC
     2941 AAGCAGTCTT TCCCAGAAGG TTTTAAATGG GAAAGGGTCA TGAACTTTGA GGATGGTGGT
     3001 GCTGTGACGG TGACGCAGGA TACTTCTTTG GAGGACGGAA CACTCATCTA CAAGGTAAAG
     3061 CTCAGGGGCA CGAACTTCCC GCCGGACGGA CCAGTCATGC AGAAAAAGAC TATGGGATGG
     3121 GAGGCTTCCA CCGAACGGTT GTATCCCGAG GACGGTGTcT TGAAGGGAGA TATAAAAATG
     3181 GCTCTTCGGC TGAAAGATGG AGGCAGATAC TTGGCAGATT TTAAGACCAC GTATAAGGCA
     3241 AAAAAACCAG TGCAAATGCC GGGAGCATAT AACGTTGACC GCAAACTGGA CATCACCTCA
     3301 CATAACGAGG ATTATACAGT gGTAGAACAG TACGAAAGAA GCGAAGGGAG GCACAGCACC
     3361 GGGGGTATGG ATGAGCTGTA TAAGggcagc ggcagcggca gcatgattac cgcgccgcgc
     3421 gaatttgatt atctgaacct gcgcgcgggc ctgagcatga cctgaACGCG TCATGCTCCG
     3481 GGGTTGGGGT TGCGCCTTTT CCAAGGCAGC CCTGGGTTTG CGCAGGGACG CGGCTGCTCT
     3541 GGGCGTGGTT CCGGGAAACG CAGCGGCGCC GACCCTGGGT CTCGCACATT CTTCACGTCC
     3601 GTTCGCAGCG TCACCCGGAT CTTCGCCGCT ACCCTTGTGG GCCCCCCGGC GACGCTTCCT
     3661 GCTCCGCCCC TAAGTCGGGA AGGTTCCTTG CGGTTCGCGG CGTGCCGGAC GTGACAAACG
     3721 GAAGCCGCAC GTCTCACTAG TACCCTCGCA GACGGACAGC GCCAGGGAGC AATGGCAGCG
     3781 CGCCGACCGC GATGGGCTGT GGCCAATAGC GGCTGCTCAG CAGGGCGCGC CGAGAGCAGC
     3841 GGCCGGGAAG GGGCGGTGCG GGAGGCGGGG TGTGGGGCGG TAGTGTGGGC CCTGTTCCTG
     3901 CCCGCGCGGT GTTCCGCATT CTGCAAGCCT CCGGAGCGCA CGTCGGCAGT CGGCTCCCTC
     3961 GTTGACCGAA TCACCGACCT CTCTCCCCAG CAATTCACCA TGTCTAGACT GGACAAGAGC
     4021 AAAGTCATAA ACGGAGCTCT GGAATTACTC AATGGTGTCG GTATCGAAGG CCTGACGACA
     4081 AGGAAACTCG CTCAAAAGCT GGGAGTTGAG CAGCCTACCC TGTACTGGCA CGTGAAGAAC
     4141 AAGCGGGCCC TGCTCGATGC CCTGCCAATC GAGATGCTGG ACAGGCATCA TACCCACTTC
     4201 TGCCCCCTGG AAGGCGAGTC ATGGCAAGAC TTTCTGCGGA ACAACGCCAA GTCATACCGC
     4261 TGTGCTCTCC TCTCACATCG CGACGGGGCT AAAGTGCATC TCGGCACCCG CCCAACAGAG
     4321 AAACAGTACG AAACCCTGGA AAATCAGCTC GCGTTCCTGT GTCAGCAAGG CTTCTCCCTG
     4381 GAGAACGCAC TGTACGCTCT GTCCGCCGTG GGCCACTTTA CACTGGGCTG CGTATTGGAG
     4441 GAACAGGAGC ATCAAGTAGC AAAAGAGGAA AGAGAGACAC CTACCACCGA TTCTATGCCC
     4501 CCACTTCTGA GACAAGCAAT TGAGCTGTTC GACCGGCAGG GAGCCGAACC TGCCTTCCTT
     4561 TTCGGCCTGG AACTAATCAT ATGTGGCCTG GAGAAACAGC TAAAGTGCGA AAGCGGCGGG
     4621 CCGACCGACG CCCTTGACGA TTTTGACTTA GACATGCTCC CAGCCGATGC CCTTGACGAC
     4681 TTTGACCTTG ATATGCTGCC TGCTGACGCT CTTGACGATT TTGACCTTGA CATGCTCCCC
     4741 GGGTAACTAA GTAAGGATCG ATCCAAggta ccTTGAGCAT CTGACTTCTG GCTAATAAAG
     4801 GAAATTTATT TTCATTGCAA TAGTGTGTTG GAATTTTTTG TGTCTCTCAC TCGGAAGGAC
     4861 ATATGGGAGG GCAAATCATT TAAAACATCA GAATGAGTAT TTGGTTTAGA GTTTGGCAAC
     4921 ATATGCCATA TGCTGGCTGC CATGAACAAA GGTGGCTATA AAGAGGTCAT CAGTATATGA
     4981 AACAGCCCCC TGCTGTCCAT TCCTTATTCC ATAGAAAAGC CTTGACTTGA GGTTAGATTT
     5041 TTTTTATATT TTGTTTTGTG TTATTTTTTT CTTTAACATC CCTAAAATTT TCCTTACATG
     5101 TTTTACTAGC CAGATTTTTC CTCCTCTCCT GACTACTCCC AGTCATAGCT GTCCCTCTTC
     5161 TCTTATGAAG ATCCCTCGAC CTGCAGCCCA AGCTTGGCGT AATCATGGTC ATAGCTGTTT
     5221 CCTGTGTGAA ATTGTTATCC GCTCACAATT CCACACAACA TACGAGCCGG AAGCATAAAG
     5281 TGTAAAGCCT GGGGTGCCTA ATGAGTGAGC TAACTCACAT TAATTGCGTT GCGCTCACTG
     5341 CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC CAGCGGATCC GCATCTCAAT TAGTCAGCAA
     5401 CCATAGTCCC GCCCCTAACT CCGCCCATCC CGCCCCTAAC TCCGCCCAGT TCCGCCCATT
     5461 C
//