LOCUS       pRC065d.g1              5653 bp ds-DNA     circular     20-MAR-2024
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    complement(22..648)
                     /label="TetR"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             complement(22..648)
                     /label="Translation 648-22"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS*"
     misc_feature    649..721
                     /label="tet promoter region"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    670..687
                     /label="Q1, Q2 promoter (tetR)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(697..746)
                     /label="oRC148"
                     /note="sequence: CAGTTTTCTCCTCTTTTAGTATATCttttctctatcactgatagggagtg"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    722..748
                     /label="bujRBS_mut"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          732..773
                     /label="oRC147"
                     /note="sequence: AAGAGGAGAAAACTGATATGatcgaaaaaaaaaagtccttcg"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             749..3649
                     /label="Translation 749-3649"
                     /translation="MIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSIRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEKRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVYTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIINYGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKDMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVINLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAGRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFFKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDENGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQKKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRENAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEEKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKAEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHAYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVPFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             752..3649
                     /label="RfxCas13d"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="product: RNA-targeting Type VI CRISPR protein from Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,2018) translation: IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSIRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEKRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVYTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIINYGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKDMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVINLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAGRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFFKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDENGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQKKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRENAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEEKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKAEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHAYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVPFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     misc_feature    1463..1465
                     /label="R239A"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    1478..1480
                     /label="H244A"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    3320..3322
                     /label="R858A"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3335..3337
                     /label="H863A"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(3633..3669)
                     /label="oRC149"
                     /note="sequence: ggagatccttactcgagttaggaattgccggacacct"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             3650..3652
                     /label="Translation 3650-3652"
                     /translation="*"
     primer          3654..3673
                     /label="oCK064"
                     /note="sequence: tcgagtaaggatctccaggc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     terminator      3668..3735
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     terminator      3668..3796
                     /label="dblTerm"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_fwdcolor="#800080"
     misc_feature    3797..4514
                     /label="p15A origin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    3858..3877
                     /label="oCD007"
                     /ApEinfo_revcolor="#004080"
                     /ApEinfo_fwdcolor="#004080"
     misc_feature    3950..4512
                     /label="p15A origin"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          4465..4484
                     /label="oCD244"
                     /note="sequence: caagagattacgcgcagacc"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(4551..4578)
                     /label="oBT055_183_factR"
                     /note="sequence: gctgacttcaggtgctacatttgaagag"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(4561..4580)
                     /label="oCD349"
                     /note="sequence: gggctgacttcaggtgctac"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          4589..4626
                     /label="oJF153"
                     /note="sequence: ATAAGTTGTTACTAGATTGACAGCTAGCTCAGTCCTAG"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     primer          4589..4648
                     /label="oRC152"
                     /note="sequence: ataagttgttactagATTGACAGCTAGCTCAGTCCTAGGTATAATACTAGTCACTAGTGC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          4605..4626
                     /label="oCK132"
                     /note="sequence: cctgTGGCAATTCCGACGTCTTGACAGCTAGCTCAGTCCTAG"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     promoter        4605..4639
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    4640..4669
                     /label="30 DR"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    4670..4697
                     /label="crRNA"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(4698..4714)
                     /label="oRC171"
                     /note="sequence: GGGCTTTctcatgcgtt"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     terminator      4698..4750
                     /label="ECK120033736 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(4726..4776)
                     /label="oCK944"
                     /note="sequence: ggtgagaatccaagcactagCTTAAGgcgcaataaaAAAGCCCCCGGAAGG"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    4757..4769
                     /label="overhang"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    4762..4769
                     /label="Phage lambda T0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    4770..4867
                     /label="Phage lambda T0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(4777..5640)
                     /label="CmR(EcoRI-KO) "
                     /ApEinfo_revcolor="#120eff"
                     /ApEinfo_fwdcolor="#120eff"
     primer          complement(4801..4821)
                     /label="oCD245"
                     /note="sequence: ctggatttgttcagaacgctc"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(4855..4872)
                     /label="oRC053"
                     /note="sequence: agctcgcttggactcctg"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(4868..4892)
                     /label="oCK446"
                     /note="sequence: cggggcgtaatttgatatcgagctc"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    complement(4886..5542)
                     /label="CmR (EcoRI-KO)"
                     /ApEinfo_revcolor="#000bff"
                     /ApEinfo_fwdcolor="#000bff"
     primer          complement(4926..4946)
                     /label="oCD491"
                     /note="sequence: CCATGTCGGCAGAATGCTTAA"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(4942..4961)
                     /label="oCK089"
                     /note="sequence: cgtttgtgatggcttccatg"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     promoter        complement(5609..5640)
                     /label="pCat promoter (weak)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
ORIGIN
        1 ttttcgccag atatcgacgt cttaagaccc actttcacat ttaagttgtt tttctaatcc
       61 gcatatgatc aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa
      121 ttcgatagct tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc
      181 tttagcgact tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc
      241 gctgagtgca tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg
      301 attttcgaga gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc
      361 atcgcgatga cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg
      421 ccagctttcc ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc
      481 taaggcgtcg agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac
      541 acctagcttc tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag
      601 ctctaatgcg ctgttaatca ctttactttt atctaatcta gacatcatta attcctaatt
      661 tttgttgaca ctctatcgtt gatagagtta ttttaccact ccctatcagt gatagagaaa
      721 aGATATACTA AAAGAGGAGA AAACTGATAT Gatcgaaaaa aaaaagtcct tcgccaaggg
      781 catgggcgtg aagtccacac tcgtgtccgg ctccaaagtg tacatgacaa ccttcgccga
      841 aggcagcgac gccaggctgg aaaagatcgt ggagggcgac agcatcagga gcgtgaatga
      901 gggcgaggcc ttcagcgctg aaatggccga taaaaacgcc ggctataaga tcggcaacgc
      961 caaattcagc catcctaagg gctacgccgt ggtggctaac aaccctctgt atacaggacc
     1021 cgtccagcag gatatgctcg gcctgaagga aactctggaa aagaggtact tcggcgagag
     1081 cgctgatggc aatgacaata tttgtatcca ggtgatccat aacatcctgg acattgaaaa
     1141 aatcctcgcc gaatacatta ccaacgccgc ctacgccgtc aacaatatct ccggcctgga
     1201 taaggacatt attggattcg gcaagttctc cacagtgtat acctacgacg aattcaaaga
     1261 ccccgagcac catagggccg ctttcaacaa taacgataag ctcatcaacg ccatcaaggc
     1321 ccagtatgac gagttcgaca acttcctcga taaccccaga ctcggctatt tcggccaggc
     1381 ctttttcagc aaggagggca gaaattacat catcaattac ggcaacgaat gctatgacat
     1441 tctggccctc ctgagcggac tggcgcactg ggtggtcgct aacaacgaag aagagtccag
     1501 gatctccagg acctggctct acaacctcga taagaacctc gacaacgaat acatctccac
     1561 cctcaactac ctctacgaca ggatcaccaa tgagctgacc aactccttct ccaagaactc
     1621 cgccgccaac gtgaactata ttgccgaaac tctgggaatc aaccctgccg aattcgccga
     1681 acaatatttc agattcagca ttatgaaaga gcagaaaaac ctcggattca atatcaccaa
     1741 gctcagggaa gtgatgctgg acaggaagga tatgtccgag atcaggaaaa atcataaggt
     1801 gttcgactcc atcaggacca aggtctacac catgatggac tttgtgattt ataggtatta
     1861 catcgaagag gatgccaagg tggctgccgc caataagtcc ctccccgata atgagaagtc
     1921 cctgagcgag aaggatatct ttgtgattaa cctgaggggc tccttcaacg acgaccagaa
     1981 ggatgccctc tactacgatg aagctaatag aatttggaga aagctcgaaa atatcatgca
     2041 caacatcaag gaatttaggg gaaacaagac aagagagtat aagaagaagg acgcccctag
     2101 actgcccaga atcctgcccg ctggccgtga tgtttccgcc ttcagcaaac tcatgtatgc
     2161 cctgaccatg ttcctggatg gcaaggagat caacgacctc ctgaccaccc tgattaataa
     2221 attcgataac atccagagct tcctgaaggt gatgcctctc atcggagtca acgctaagtt
     2281 cgtggaggaa tacgcctttt tcaaagactc cgccaagatc gccgatgagc tgaggctgat
     2341 caagtccttc gctagaatgg gagaacctat tgccgatgcc aggagggcca tgtatatcga
     2401 cgccatccgt attttaggaa ccaacctgtc ctatgatgag ctcaaggccc tcgccgacac
     2461 cttttccctg gacgagaacg gaaacaagct caagaaaggc aagcacggca tgagaaattt
     2521 cattattaat aacgtgatca gcaataaaag gttccactac ctgatcagat acggtgatcc
     2581 tgcccacctc catgagatcg ccaaaaacga ggccgtggtg aagttcgtgc tcggcaggat
     2641 cgctgacatc cagaaaaaac agggccagaa cggcaagaac cagatcgaca ggtactacga
     2701 aacttgtatc ggaaaggata agggcaagag cgtgagcgaa aaggtggacg ctctcacaaa
     2761 gatcatcacc ggaatgaact acgaccaatt cgacaagaaa aggagcgtca ttgaggacac
     2821 cggcagggaa aacgccgaga gggagaagtt taaaaagatc atcagcctgt acctcaccgt
     2881 gatctaccac atcctcaaga atattgtcaa tatcaacgcc aggtacgtca tcggattcca
     2941 ttgcgtcgag cgtgatgctc aactgtacaa ggagaaaggc tacgacatca atctcaagaa
     3001 actggaagag aagggattca gctccgtcac caagctctgc gctggcattg atgaaactgc
     3061 ccccgataag agaaaggacg tggaaaagga gatggctgaa agagccaagg agagcattga
     3121 cagcctcgag agcgccaacc ccaagctgta tgccaattac atcaaataca gcgacgagaa
     3181 gaaagccgag gagttcacca ggcagattaa cagggagaag gccaaaaccg ccctgaacgc
     3241 ctacctgagg aacaccaagt ggaatgtgat catcagggag gacctcctga gaattgacaa
     3301 caagacatgt accctgttcg caaacaaggc cgtcgccctg gaagtggcca ggtatgtcca
     3361 cgcctatatc aacgacattg ccgaggtcaa ttcctacttc caactgtacc attacatcat
     3421 gcagagaatt atcatgaatg agaggtacga gaaaagcagc ggaaaggtgt ccgagtactt
     3481 cgacgctgtg aatgacgaga agaagtacaa cgataggctc ctgaaactgc tgtgtgtgcc
     3541 tttcggctac tgtatcccca ggtttaagaa cctgagcatc gaggccctgt tcgataggaa
     3601 cgaggccgcc aagttcgaca aggagaaaaa gaaggtgtcc ggcaattcct aactcgagta
     3661 aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     3721 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     3781 cctttctgcg tttataccta gggatatatt ccgcttcctc gctcactgac tcgctacgct
     3841 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     3901 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     3961 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     4081 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     4141 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     4201 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     4261 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     4321 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     4381 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     4441 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     4501 gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
     4561 gtagcacctg aagtcagccc catacgatat aagttgttac tagATTGACA GCTAGCTCAG
     4621 TCCTAGGTAT AATACTAGTC ACTAGTGCGA ATTTGCACTA GTCTAAAACT GGTACCTTTC
     4681 TCCTCTTTAA TGAATTCaac gcatgagAAA GCCCCCGGAA GATCACCTTC CGGGGGCTTT
     4741 tttattgcgc CTTAAGctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     4801 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     4861 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     4921 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     4981 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     5041 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     5101 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     5161 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     5221 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     5281 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     5341 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     5401 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     5461 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     5521 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5581 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5641 gatcaacgtc tca
//