LOCUS       pZH521_const_Tet        4403 bp ds-DNA     circular     26-SEP-2017
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     pZH521 -- modified pZH509 to have constitutive TetR expression
            w/out bicistronic expression and low GFPmut2 RBS strength
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     promoter        189..262
                     /locus_tag="pLtetO-1"
                     /ApEinfo_label="pLtetO-1"
                     /ApEinfo_fwdcolor="#18ff11"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     gene            3325..4403
                     /locus_tag="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    189..207
                     /locus_tag="TetO2"
                     /ApEinfo_label="TetO2"
                     /ApEinfo_fwdcolor="#ff120b"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    214..232
                     /locus_tag="TetO2(1)"
                     /ApEinfo_label="TetO2"
                     /ApEinfo_fwdcolor="#ff120b"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      1891..1934
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYKRPAANDENYAASV*"
                     /locus_tag="rrnB_T1_terminator"
                     /ApEinfo_label="rrnB_T1_terminator"
                     /ApEinfo_fwdcolor="#b1a0ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      1018..1061
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYKRPAANDENYAASV*"
                     /locus_tag="rrnB_T1_terminator(1)"
                     /ApEinfo_label="rrnB_T1_terminator"
                     /ApEinfo_fwdcolor="#b1a0ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     RBS             1206..1211
                     /locus_tag="RBS"
                     /ApEinfo_label="RBS"
                     /ApEinfo_fwdcolor="#80ff80"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      2340..3183
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
                     SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
                     ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
                     A*"
                     /locus_tag="p15A_origin"
                     /ApEinfo_label="p15A_origin"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     -35_signal      208..213
                     /locus_tag="-33"
                     /ApEinfo_label="-33"
                     /ApEinfo_fwdcolor="#ff4669"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     -10_signal      231..236
                     /locus_tag="-10"
                     /ApEinfo_label="-10"
                     /ApEinfo_fwdcolor="#ff5d86"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    243..243
                     /locus_tag="pLtetO-1 +1"
                     /ApEinfo_label="pLtetO-1 +1"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1220..1843
                     /locus_tag="TetR tn10"
                     /ApEinfo_label="TetR tn10"
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             292..1005
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITHGMDELYK"
                     /locus_tag="GFPmut2"
                     /ApEinfo_label="GFPmut2"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        1062..1205
                     /locus_tag="proB insulated promoter"
                     /ApEinfo_label="proB insulated promoter"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     -35_signal      1124..1129
                     /locus_tag="ProB/D -35"
                     /ApEinfo_label="ProB/D -35"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     -10_signal      1147..1152
                     /locus_tag="ProB -10"
                     /ApEinfo_label="ProB -10"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1159..1159
                     /locus_tag="ProD +1"
                     /ApEinfo_label="ProD +1"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA
       61 CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG
      121 TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC
      181 ACCATATGTC CCTATCAGTG ATAGAGATTG ACATCCCTAT CAGTGATAGA GATACTGAGC
      241 ACATCAGCAG GACGCACTGA CCgaattcat taaaTTCgag aaaggtaccg catgagtaaa
      301 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
      361 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
      421 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
      481 ttcgcgtatg gtcttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt
      541 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
      601 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
      661 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
      721 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
      781 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
      841 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
      901 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
      961 gtaacagctg ctgggattac acatggcatg gatgaactat acaaaTAAta atctagcata
     1021 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tCACAGCTAA CACCACGTCG
     1081 TCCCTATCTG CTGCCCTAGG TCTATGAGTG GTTGCTGGAT AACTTTACGG GCATGCATAA
     1141 GGCTCGTAat ATATATTCAG GGAGACCACA ACGGTTTCCC TCTACAAATA ATTTTGTTTA
     1201 ACTTTaggag gatttcaccA TGTCTAGATT AGATAAAAGT AAAGTGATTA ACAGCGCATT
     1261 AGAGCTGCTT AATGAGGTCG GAATCGAAGG TTTAACAACC CGTAAACTCG CCCAGAAGCT
     1321 AGGTGTAGAG CAGCCTACAT TGTATTGGCA TGTAAAAAAT AAGCGGGCTT TGCTCGACGC
     1381 CTTAGCCATT GAGATGTTAG ATAGGCACCA TACTCACTTT TGCCCTTTAG AAGGGGAAAG
     1441 CTGGCAAGAT TTTTTACGTA ATAACGCTAA AAGTTTTAGA TGTGCTTTAC TAAGTCATCG
     1501 CGATGGAGCA AAAGTACATT TAGGTACACG GCCTACAGAA AAACAGTATG AAACTCTCGA
     1561 AAATCAATTA GCCTTTTTAT GCCAACAAGG TTTTTCACTA GAGAATGCAT TATATGCACT
     1621 CAGCGCTGTG GGGCATTTTA CTTTAGGTTG CGTATTGGAA GATCAAGAGC ATCAAGTCGC
     1681 TAAAGAAGAA AGGGAAACAC CTACTACTGA TAGTATGCCG CCATTATTAC GACAAGCTAT
     1741 CGAATTATTT GATCACCAAG GTGCAGAGCC AGCCTTCTTA TTCGGCCTTG AATTGATCAT
     1801 CTGCGGATTA GAAAAACAAC TTAAATGTGA AAGTGGGTCT TAATAActgc agcccggggg
     1861 atcccatggt acgcgtgcta gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1921 gcctttcgtt ttatAAGCTT GGCGTAATCA TGGTCATAGC TGTTTCCTGT GTGAAATTGT
     1981 TATCCGCTCA CAATTCCACA CAACATACGA GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT
     2041 GCCTAATGAG TGAGCTAACT CACATTAATT GCGTTGCGCT CACTGCCCGC TTTCCAGTCG
     2101 GGAAACCTGT CGTGCCAGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG
     2161 CGTATTGGGC GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG
     2221 CGGCGAGCGG TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT
     2281 AACGCAGGAA AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCg
     2341 cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc
     2401 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat
     2461 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     2521 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     2581 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     2641 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     2701 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc
     2761 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc
     2821 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt
     2881 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc
     2941 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa
     3001 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt
     3061 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc
     3121 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat
     3181 attGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT
     3241 GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT
     3301 TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG
     3361 TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT
     3421 CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC
     3481 GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC
     3541 CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG
     3601 GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC
     3661 AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG
     3721 ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC
     3781 TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT
     3841 GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC
     3901 AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT
     3961 ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC
     4021 TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC
     4081 TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA
     4141 AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT
     4201 CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG
     4261 ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG
     4321 AAAAGTGCCA CCTGACGTCT AAGAAACCAT TATTATCATG ACATTAACCT ATAAAAATAG
     4381 GCGTATCACG AGGCCCTTTC GTC   
//
