LOCUS       pSECRETS_A              6948 bp ds-DNA     circular     17-JAN-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     
COMMENT     ApEinfo:methylated:0
FEATURES             Location/Qualifiers
     misc_feature    631..735
                     /locus_tag="Amp promotor"
                     /label="Amp promotor"
                     /ApEinfo_label="Amp promotor"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    538..630
                     /locus_tag="tetR"
                     /label="tetR"
                     /ApEinfo_label="tetR"
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    812..885
                     /locus_tag="pltetO1 promoter"
                     /label="pltetO1 promoter"
                     /ApEinfo_label="pltetO1 promoter"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      5113..5184
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /locus_tag="rrnB T1 terminator"
                     /label="rrnB T1 terminator"
                     /ApEinfo_label="rrnB T1 terminator"
                     /ApEinfo_fwdcolor="#9d1b1c"
                     /ApEinfo_revcolor="#9d1b1c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(5255..5272)
                     /note="L4440 vector, forward primer"
                     /locus_tag="L4440"
                     /label="L4440"
                     /ApEinfo_label="L4440"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(5389..5934)
                     /direction=LEFT
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin. "
                     /locus_tag="p15A ori"
                     /label="p15A ori"
                     /ApEinfo_label="p15A ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(5469..5488)
                     /note="pBR322 origin, forward primer"
                     /locus_tag="pBR322ori-F"
                     /label="pBR322ori-F"
                     /ApEinfo_label="pBR322ori-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      6048..6142
                     /note="transcription terminator from phage lambda"
                     /locus_tag="lambda t0 terminator"
                     /label="lambda t0 terminator"
                     /ApEinfo_label="lambda t0 terminator"
                     /ApEinfo_fwdcolor="#9d1b1c"
                     /ApEinfo_revcolor="#9d1b1c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(6163..6822)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     L
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS
                     S
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN
                     M
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
                     /locus_tag="CmR"
                     /label="CmR"
                     /ApEinfo_label="CmR"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     6737..6756
                     /note="Chloramphenicol resistance gene, reverse primer"
                     /locus_tag="CAT-R"
                     /label="CAT-R"
                     /ApEinfo_label="CAT-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(6823..6925)
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
                     /locus_tag="cat promoter"
                     /label="cat promoter"
                     /ApEinfo_label="cat promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(7..537)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /note="TetR binds to the tetracycline operator tetO to
                     inhibit transcription. This inhibition can be relieved by
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     V
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     R
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     T DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
                     /locus_tag="TetR"
                     /label="TetR"
                     /ApEinfo_label="TetR"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    5200..5227
                     /locus_tag="T7Te terminator"
                     /label="T7Te terminator"
                     /ApEinfo_label="T7Te terminator"
                     /ApEinfo_fwdcolor="#800000"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    983..5089
                     /locus_tag="Cas9"
                     /label="Cas9"
                     /ApEinfo_label="Cas9"
                     /ApEinfo_fwdcolor="#ffff00"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgGGC
      541 GAGTTTACGG GTTGTTAaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta aacgagacat actcttcctt tttcaatatt attgaagcat
      661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcctct ggctaactag
      781 agaacccact gcttactggc ttatcgaaat ttccctatca gtgatagaga ttgacatccc
      841 tatcagtgat agagatactg agcacatcag caggacgcac tgaccaggga gacccaagct
      901 tgccaccatg gtgtacccct acgacgtgcc cgactacgcc gaattgcctc caaaaaagaa
      961 gagaaaggta gggatccgaa ccatggataa gaaatactca ataggcttag atatcgGCAC
     1021 AAATAGCGTC GGATGGgcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa
     1081 ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt
     1141 tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac
     1201 acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt
     1261 agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca
     1321 tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc
     1381 aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt
     1441 aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga
     1501 tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa
     1561 tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc
     1621 tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa
     1681 gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa
     1741 atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga
     1801 tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc
     1861 taagaattta tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac
     1921 taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac
     1981 tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga
     2041 tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag aagaatttta
     2101 taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact
     2161 aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca
     2221 aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt
     2281 aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg
     2341 tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat
     2401 taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga
     2461 acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt
     2521 gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg
     2581 aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt
     2641 caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga
     2701 atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac
     2761 ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga
     2821 agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga
     2881 ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg
     2941 tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa
     3001 gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt
     3061 tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt
     3121 gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat
     3181 taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg
     3241 gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg
     3301 ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag
     3361 tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct
     3421 ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt
     3481 aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg attcaataga
     3541 caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga
     3601 agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac
     3661 tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa
     3721 agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca
     3781 aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt
     3841 taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta
     3901 taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt
     3961 tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta
     4021 taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac
     4081 cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc
     4141 aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt
     4201 ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa
     4261 tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa
     4321 aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg
     4381 ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa
     4441 atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc
     4501 ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga
     4561 cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat
     4621 gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt
     4681 gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga
     4741 acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat
     4801 cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc
     4861 atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt
     4921 tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg
     4981 taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac
     5041 tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactaac tcgagtaagg
     5101 atctccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5161 tgttgtttgt cggtgaacgc tctctactag AGTCACACTG GCTCACCTTC gggtgggcct
     5221 ttctgcgttt atacctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg
     5281 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     5341 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     5401 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     5461 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     5521 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     5581 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     5641 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     5701 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     5761 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     5821 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     5881 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     5941 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     6001 gcacctgaag tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata
     6061 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     6121 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
     6181 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     6241 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     6301 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     6361 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     6421 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     6481 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     6541 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     6601 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     6661 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     6721 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     6781 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     6841 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     6901 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc cagatatc 
//
