LOCUS       MS2_SC2_S_Kappa         6264 bp ds-DNA     circular     23-AUG-2021
DEFINITION  .
KEYWORDS    "Antibiotic Resistance"                                            
FEATURES             Location/Qualifiers
     promoter        3..32
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     RBS             52..74
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             131..1312
                     /label="Maturation Protein"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    166..166
                     /label="g->a (remove BsaI)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    715..715
                     /label="g->a (remove BspMI)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    961..961
                     /label="t->c (remove BsmBI)"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    1336..1725
                     /label="Coat Protein"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    1336..2136
                     /label="coat protein dimer"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    1671..1671
                     /label="c->t (remove BsaI)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    1729..1731
                     /label="Start of second coat dimer"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1768..1785
                     /label="His Tag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    2079..2079
                     /label="c->t (remove BsaI)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     terminator      2150..2211
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          2210..2237
                     /label="pACYC_MS2_PLP_RFPpac FWD (1)"
                     /note="sequence: tgTCGAACAGAAAGTAATCGTATTGTAC"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    2212..2256
                     /label="Spacer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          2276..2301
                     /label="MS2 SC2 Variant GG rev"
                     /note="sequence: TCACGTCTCAATAGGAGAGTCCAACCAACAGAATCTATTGT"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    2276..3929
                     /label="insert"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    2675..2677
                     /label="L452R"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    2771..2773
                     /label="E484Q"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    3362..3364
                     /label="P681R"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(3905..3929)
                     /label="MS2 SC2 Variant GG fwd"
                     /note="sequence: CCTCGTCTCCATGTCATTTCATCTGTGAGCAAAGGTGG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     terminator      3948..4009
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(4037..4056)
                     /label="MS2 IC RNA Rev"
                     /note="sequence: CAGCTGGCACGACAGGTTTC"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          complement(4063..4084)
                     /label="circRNA rev"
                     /note="sequence: CAAGGAAACAGCTATGACCATG"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(4066..4082)
                     /label="[Eurofins] M13 rev (-29)"
                     /note="sequence: CAGGAAACAGCTATGACC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     rep_origin      complement(4199..4881)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          complement(4204..4225)
                     /label="HIV Seq Fwd"
                     /note="sequence: ctatggaaaaacgccagcaacg"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     CDS             complement(4976..5773)
                     /label="KanR"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     promoter        6215..6233
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     protein_bind    6234..6258
                     /label="lac operator"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
ORIGIN
        1 gatttacact ttatgcttcc ggctcgtatg tttctagaga attcaaataa ttttgtttaa
       61 ctttaagaag gagatttaaa tTGGCTATCG CTGTAGGTAG CCGGAATTCC ATTCCTAGGA
      121 GGTTTGACCT gtgcgagctt ttagtaccct tgatagggag aacgaaacct tcgtcccctc
      181 cgttcgcgtt tacgcggacg gtgagactga agataactca ttctctttaa aatatcgttc
      241 gaactggact cccggtcgtt ttaactcgac tggggccaaa acgaaacagt ggcactaccc
      301 ctctccgtat tcacgggggg cgttaagtgt cacatcgata gatcaaggtg cctacaagcg
      361 aagtgggtca tcgtggggtc gcccgtacga ggagaaagcc ggtttcggct tctccctcga
      421 cgcacgctcc tgctacagcc tcttccctgt aagccaaaac ttgacttaca tcgaagtgcc
      481 gcagaacgtt gcgaaccggg cgtcgaccga agtcctgcaa aaggtcaccc agggtaattt
      541 taaccttggt gttgctttag cagaggccag gtcgacagcc tcacaactcg cgacgcaaac
      601 cattgcgctc gtgaaggcgt acactgccgc tcgtcgcggt aattggcgcc aggcgctccg
      661 ctaccttgcc ctaaacgaag atcgaaagtt tcgatcaaaa cacgtggccg gcagatggtt
      721 ggagttgcag ttcggttggt taccactaat gagtgatatc cagggtgcat atgagatgct
      781 tacgaaggtt caccttcaag agtttcttcc tatgagagcc gtacgtcagg tcggtactaa
      841 catcaagtta gatggccgtc tgtcgtatcc agctgcaaac ttccagacaa cgtgcaacat
      901 atcgcgacgt atcgtgatat ggttttacat aaacgatgca cgtttggcat ggttgtcgtc
      961 cctaggtatc ttgaacccac taggtatagt gtgggaaaag gtgcctttct cattcgttgt
     1021 cgactggctc ctacctgtag gtaacatgct cgagggcctt acggcccccg tgggatgctc
     1081 ctacatgtca ggaacagtta ctgacgtaat aacgggtgag tccatcataa gcgttgacgc
     1141 tccctacggg tggactgtgg agagacaggg cactgctaag gcccaaatct cagccatgca
     1201 tcgaggggta caatccgtat ggccaacaac tggcgcgtac gtaaagtctc ctttctcgat
     1261 ggtccatacc ttagatgcgt tagcattaat caggcaacgg ctctctagat agAGCCCTCA
     1321 ACCGGAGTTT GAAGCATGGC TTCTAACTTT ACTCAGTTCG TTCTCGTCGA CAATGGCGGA
     1381 ACTGGCGACG TGACTGTCGC CCCAAGCAAC TTCGCTAACG GGGTCGCTGA ATGGATCAGC
     1441 TCTAACTCGC GTTCACAGGC TTACAAAGTA ACCTGTAGCG TTCGTCAGAG CTCTGCGCAG
     1501 AATCGCAAAT ACACCATCAA AGTCGAGGTG CCTAAAGTGG CAACCCAGAC TGTTGGTGGT
     1561 GTAGAGCTTC CTGTAGCCGC ATGGCGTTCG TACTTAAATA TGGAACTAAC CATTCCAATT
     1621 TTCGCTACGA ATTCCGACTG CGAGCTTATT GTTAAGGCAA TGCAAGGTCT TCTAAAAGAT
     1681 GGAAACCCGA TTCCCTCAGC AATCGCAGCA AACTCCGGCA TCTACGCTAA CTTTACTCAG
     1741 TTCGTTCTCG TCGACAATGG CGGTACCCAT CACCATCACC ATCACGGTAC CGGCGACGTG
     1801 ACTGTCGCCC CAAGCAACTT CGCTAACGGG GTCGCTGAAT GGATCAGCTC TAACTCGCGT
     1861 TCACAGGCTT ACAAAGTAAC CTGTAGCGTt CGTCAGAGCT CTGCGCAGAA TCGCAAATAC
     1921 ACCATCAAAG TCGAGGTGCC TAAAGTGGCA ACCCAGACTG TTGGTGGTGT AGAGCTTCCT
     1981 GTAGCCGCAT GGCGTTCGTA CTTAAATATG GAACTAACCA TTCCAATTTT CGCTACGAAT
     2041 TCCGACTGCG AGCTTATTGT TAAGGCAATG CAAGGTCTTC TAAAAGATGG AAACCCGATT
     2101 CCCTCAGCAA TCGCAGCAAA CTCCGGCATC TACTAATAGA CGCCGGCCAc cgctgagcaa
     2161 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gTCGAACAGA
     2221 AAGTAATCGT ATTGTACACG GCCGCATAAT CGAAATTAAT ACGACTCACT ATAGGagagt
     2281 ccaaccaaca gaatctattg ttagatttcc taatattaca aacttgtgcc cttttggtga
     2341 agtttttaac gccaccagat ttgcatctgt ttatgcttgg aacaggaaga gaatcagcaa
     2401 ctgtgttgct gattattctg tcctatataa ttccgcatca ttttccactt ttaagtgtta
     2461 tggagtgtct cctactaaat taaatgatct ctgctttact aatgtctatg cagattcatt
     2521 tgtaattaga ggtgatgaag tcagacaaat cgctccaggg caaactggaa agattgctga
     2581 ttataattat aaattaccag atgattttac aggctgcgtt atagcttgga attctaacaa
     2641 tcttgattct aaggttggtg gtaattataa ttaccggtat agattgttta ggaagtctaa
     2701 tctcaaacct tttgagagag atatttcaac tgaaatctat caggccggta gcacaccttg
     2761 taatggtgtt caaggtttta attgttactt tcctttacaa tcatatggtt tccaacccac
     2821 taatggtgtt ggttaccaac catacagagt agtagtactt tcttttgaac ttctacatgc
     2881 accagcaact gtttgtggac ctaaaaagtc tactaatttg gttaaaaaca aatgtgtcaa
     2941 tttcaacttc aatggtttaa caggcacagg tgttcttact gagtctaaca aaaagtttct
     3001 gcctttccaa caatttggca gagacattgc tgacactact gatgctgtcc gtgatccaca
     3061 gacacttgag attcttgaca ttacaccatg ttcttttggt ggtgtcagtg ttataacacc
     3121 aggaacaaat acttctaacc aggttgctgt tctttatcag gatgttaact gcacagaagt
     3181 ccctgttgct attcatgcag atcaacttac tcctacttgg cgtgtttatt ctacaggttc
     3241 taatgttttt caaacacgtg caggctgttt aataggggct gaacatgtca acaactcata
     3301 tgagtgtgac atacccattg gtgcaggtat atgcgctagt tatcagactc agactaattc
     3361 tcgtcggcgg gcacgtagtg tagctagtca atccatcatt gcctacacta tgtcacttgg
     3421 tgcagaaaat tcagttgctt actctaataa ctctattgcc atacccacaa attttactat
     3481 tagtgttacc acagaaattc taccagtgtc tatgaccaag acatcagtag attgtacaat
     3541 gtacatttgt ggtgattcaa ctgaatgcag caatcttttg ttgcaatatg gcagtttttg
     3601 tacacaatta aaccgtgctt taactggaat agctgttgaa caagacaaaa acacccaaga
     3661 agtttttgca caagtcaaac aaatttacaa aacaccacca attaaagatt ttggtggttt
     3721 taatttttca caaatattac cagatccatc aaaaccaagc aagaggtcat ttattgaaga
     3781 tctacttttc aacaaagtga cacttgcaga tgctggcttc atcaaacaat atggtgattg
     3841 ccttggtgat attgctgcta gagacctcat ttgtgcacaa aagtttaacg gccttactgt
     3901 tttgccacct ttgctcacag atgaaatgaC ATGAGGATCA CCCATGTccg ctgagcaata
     3961 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgT AATTTCACTG
     4021 CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC CAGCTGCATT AACATGGTCA TAGCTGTTTC
     4081 CTTGCGTATT GGGCGCTCTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG
     4141 GTAAAGCCTG GGGTGCCTAA TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG
     4201 CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC
     4261 GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG
     4321 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT
     4381 TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG
     4441 TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT
     4501 GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC
     4561 TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT
     4621 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC
     4681 TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA
     4741 CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT
     4801 CTCAAGAAGA TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC
     4861 GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT
     4921 AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTATT
     4981 AGAAAAATTC ATCCAGCAGA CGATAAAACG CAATACGCTG GCTATCCGGT GCCGCAATGC
     5041 CATACAGCAC CAGAAAACGA TCCGCCCATT CGCCGCCCAG TTCTTCCGCA ATATCACGGG
     5101 TGGCCAGCGC AATATCCTGA TAACGATCCG CCACGCCCAG ACGGCCGCAA TCAATAAAGC
     5161 CGCTAAAACG GCCATTTTCC ACCATAATGT TCGGCAGGCA CGCATCACCA TGGGTCACCA
     5221 CCAGATCTTC GCCATCCGGC ATGCTCGCTT TCAGACGCGC AAACAGCTCT GCCGGTGCCA
     5281 GGCCCTGATG TTCTTCATCC AGATCATCCT GATCCACCAG GCCCGCTTCC ATACGGGTAC
     5341 GCGCACGTTC AATACGATGT TTCGCCTGAT GATCAAACGG ACAGGTCGCC GGGTCCAGGG
     5401 TATGCAGACG ACGCATGGCA TCCGCCATAA TGCTCACTTT TTCTGCCGGC GCCAGATGGC
     5461 TAGACAGCAG ATCCTGACCC GGCACTTCGC CCAGCAGCAG CCAATCACGG CCCGCTTCGG
     5521 TCACCACATC CAGCACCGCC GCACACGGAA CACCGGTGGT GGCCAGCCAG CTCAGACGCG
     5581 CCGCTTCATC CTGCAGCTCG TTCAGCGCAC CGCTCAGATC GGTTTTCACA AACAGCACCG
     5641 GACGACCCTG CGCGCTCAGA CGAAACACCG CCGCATCAGA GCAGCCAATG GTCTGCTGCG
     5701 CCCAATCATA GCCAAACAGA CGTTCCACCC ACGCTGCCGG GCTACCCGCA TGCAGGCCAT
     5761 CCTGTTCAAT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC
     5821 TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA
     5881 CATTTCCCCG AAAAGTGCCA CCTAAATTGT AAGCGTTAAT ATTTTGTTAA AATTCGCGTT
     5941 AAATTTTTGT TAAATCAGCT CATTTTTTAA CCAATAGGCC GAAATCGGCA AAATCCCTTA
     6001 TAAATCAAAA GAATAGACCG AGATAGGGTT GAGTGGCCGC TACAGGGCGC TCCCATTCGC
     6061 CATTCAGGCT GCGCAACTGT TGGGAAGGGC GTTTCGGTGC GGGCCTCTTC GCTATTACGC
     6121 CAGCTGGCGA AAGGGGGATG TGCTGCAAGG CGATTAAGTT GGGTAACGCC AGGGTTTTCC
     6181 CAGTCACGAC GTTGTAAAAC GACGGCCAGT GAGGtaatac gactcactat aggggaattg
     6241 tgagcggata acaattcccc tcta
//