LOCUS       pTHS                    4295 bp ds-DNA     circular     05-JUL-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    1713..2005
                     /label="Block D"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    2832..2835
                     /label="scar 7"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    305..308
                     /label="Scar B1"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    1530..1579
                     /label="spacer"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(2811..2819)
                     /label="Lower stem 2*"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    2006..2009
                     /label="Scar 4"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    1915..1944
                     /label="30 bp spacer"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    1580..1583
                     /label="Scar 1"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(2802..2807)
                     /label="Upper stem 2*"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(1448..1475)
                     /label="Lac2  RBS"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    complement(2790..2801)
                     /label="Loop"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    2044..2047
                     /label="scar 5"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    complement(1945..2005)
                     /label="L3S2P21 Terminator"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    1584..1639
                     /label="Ptac"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(2790..2797)
                     /label="RBS"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    353..364
                     /label="spacer"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(2784..2789)
                     /label="Upper stem 2"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    2993..3612
                     /label="PBR322 ORI"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    309..1579
                     /label="Block A"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    1584..1643
                     /label="Block B"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(2781..2783)
                     /label="Start2"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    309..352
                     /label="ECK120017009 Terminator"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    complement(2772..2780)
                     /label="Lower stem 2"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    2048..2765
                     /label="Block F"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    1644..1708
                     /label="sRNA THS input A"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    1640..1643
                     /label="scar 2"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    2770..2831
                     /label="Block G"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(365..1447)
                     /label="LacI AM  (From Marionette)"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    join(3718..4295,1..238)
                     /label="Kanamycin resistance"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    2766..2769
                     /label="Scar 6"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    complement(2043..2047)
                     /label="Gly-Gly"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(2010..2043)
                     /label="Block E"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    1644..1708
                     /label="Block C"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    complement(2766..2769)
                     /label="Block G"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    2836..2891
                     /label="Block H"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     source          complement(2766..2768)
                     /label="source"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    2010..2039
                     /label="random seq"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    1709..1712
                     /label="scar 3"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(2763..2765)
                     /label="mEGFP (E.coli K12 codon optimised)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    complement(1476..1529)
                     /label=" PLacIR promoter"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(2836..2891)
                     /label="Ptac"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(2049..2762)
                     /label="mEGFP (E.coli K12 codon optimised)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    complement(2048..2048)
                     /label="Gly-Gly"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    2892..2895
                     /label="B2-scar"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    1713..1914
                     /label="TrrnB E.Coli- (T1 &T2 regions)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    complement(2802..2831)
                     /label="Toehold switch B*"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    1676..1705
                     /label="Switch B - complementary region"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    2770..2831
                     /label="Toehold Switch B"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(2040..2042)
                     /label="stop codon"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
ORIGIN
        1 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
       61 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
      121 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
      181 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga
      241 attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
      301 cgcgGTGAAG TGAAAAGAAA AAAGGCCGCA GAGCGGCCTT TTTAGTTAGA TCGGACCCCA
      361 CATCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC CAGCTGCATT AATGAATCGG
      421 CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCCAG GGTGGTTTTT CTTTTCACCA
      481 GTGAGACTGG CAACAGCTGA TTGCCCTTCA CCGCCTGGCC CTGAGAGAGT TGCAGCAAGC
      541 GGTCCACGCT GGTTTGCCCC AGCAGGCGAA AATCCTGTTT GATGGTGGTT AACGGCGGGA
      601 TATAACATGA GCTATCTTCG GTATCGTCGT ATCCCACTAC CGAGATATCC GCACCAACGC
      661 GCAGCCCGGA CTCGGTAATG GCGCGCATTG CGCCCAGCGC CATCTGATCG TTGGCAACCA
      721 GCATCGCAGT GGGAACGATG CCCTCATTCA GCATTTGCAT GGTTTGTTGA AAACCGGACA
      781 TGGCACTCCA GTCGCCTTCC CGTTCCGCTA TCGGCTGAAT TTGATTGCGA GTGAGATATT
      841 TATGCCAGCC AGCCAGACGC AGACGCGCCG AGACAGAACT TAATGGGCCC GCTAACAGCG
      901 CGATTTGCTG GTGACCCAAT GCGACCAGAT GCTCCACGCC CAGTCGCGTA CCGTCCTCAT
      961 GGGAGTAAAT AATACTGTTG ATGGGTGTCT GGTCAGAGAC ATCAAGAAAT AACGCCGGAA
     1021 CATTAGTGCA GGCAGCTTCC ACAGCAATGG CATCCTGGTC ATCCAGCGGA TAGTTAATGA
     1081 TCAGCCCACT GACGCGTTGC GCGAGAAGAT TGTGCACCGC CGCTTTACAG GCTTCGACGC
     1141 CGCTTCGTTC TACCATCGAC ACCACCACGC TGGCACCCAG TTGATCGGCG CGAGATTTAA
     1201 TCGCCGCGAC AATTTGCGAC GGCGCGTGCA GGGCCAGACT GGAGGTGGCA ACGCCAATCA
     1261 GCAACGACTG TTTGCCCGCC AGTTGTTGTG CCACGCGGTT GGGAATGTAA TTCAGCTCCA
     1321 CCATCGCCGC TTCCACTTTT TCCCGCGTTT TCGCAGAAAC GTGGCTGGCC TGGTTCACCA
     1381 CGCGGGAAAC GGTCATATAA GAGACACCGG CATACTCTGC GACATCGTAT AACGTTACTG
     1441 GTTTCATATT CACCTCCATG AATTGACTCT CTTCCGGGCG CTATCATGCC ATACCGCGAA
     1501 AGGTTTTTCA CCATTCGATG GCGCGCCGCG GCTGGGAATC TGAGACATGA GTCAAGATAT
     1561 TTGCTCGGTA ACGTATGCTG GAGTGTTGAC AATTAATCAT CGGCTCGTAT AATGTGTGGA
     1621 ATTGTGAGCG CTCACAATTA GGTGGGACCT ATTGGACCCG TTTCCAATAG GTGAACAAGA
     1681 CGATAGAACA AGCATTTGCA CTTATAGAGC TTATAAAACG AAAGGCTCAG TCGAAAGACT
     1741 GGGCCTTTCG TTTTATCTGT TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC
     1801 CGGGAGCGGA TTTGAACGTT GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC
     1861 CATAAACTGC CAGGCATCAA ATTAAGCAGA AGGCCATCCT GACGGATGGC CTTTTGGCCC
     1921 CCGACCCTTA GACTCTGTAC TCAGGGACCA AAACGAAAAA AGGCCCCCCT TTCGGGAGGC
     1981 CTCTTTTCTG GAATTTGGTA CCGAGCGCTA GTTGTATCGT ACGTCGGTCT AGGAATCGTT
     2041 TAGCCGCCTC ACTTATACAA CTCATCCATG CCTAAAGTAA TCCCCGCCGC GGTTACGAAC
     2101 TCCAAAAGGA CCATGTGATC ACGTTTCTCA TTTGGGTCCT TCGATAATTT GGATTGTGTA
     2161 CTCAGGTAGT GATTGTCAGG AAGTAAAACC GGGCCGTCGC CAATGGGTGT GTTTTGTTGA
     2221 TAATGGTCAG CCAATTGCAC GCTGCCATCC TCAATGTTGT GACGAATCTT GAAATTTACC
     2281 TTAATCCCGT TCTTCTGCTT GTCAGCCATG ATATACACGT TGTGCGAATT ATAGTTATAC
     2341 TCCAGTTTAT GTCCCAAAAT GTTGCCATCT TCTTTAAAGT CAATGCCTTT CAATTCAATG
     2401 CGATTCACTA ACGTATCCCC CTCAAACTTG ACCTCGGCGC GTGTCTTATA GTTCCCGTCA
     2461 TCCTTGAAGA AGATTGTGCG CTCTTGCACA TAGCCTTCTG GCATTGCAGA TTTAAAGAAG
     2521 TCGTGCTGCT TCATGTGATC TGGGTAACGC GAGAAACATT GGACCCCGTA TGTAAGTGTT
     2581 GTAACCAGAG TTGGCCAGGG GACTGGAAGT TTTCCTGTAG TACAAATGAA CTTAAGCGTC
     2641 AATTTGCCGT AAGTAGCGTC GCCTTCGCCC TCGCCGCTAA CCGAAAACTT ATGACCATTA
     2701 ACGTCCCCAT CCAACTCAAC CAAGATGGGT ACAACGCCTG TAAACAGCTC CTCTCCTTTA
     2761 CTCACcatCa cGCTTGTTCT CATGTCTTGT CTCCTCTGTT TCAAGACGAT AGAACAAGCA
     2821 TTTGCACTTA TtactAATTG TGAGCGCTCA CAATTCCACA CATTATACGA GCCGATGATT
     2881 AATTGTCAAC ATGCCcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2941 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3421 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3541 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3661 ttttggtcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     3721 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct
     3781 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     3841 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     3901 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     3961 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     4021 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     4081 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     4141 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     4201 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     4261 atgcataaac ttttgccatt ctcaccggat tcagt
//