LOCUS       pMC_MS2_DNA_VLP         4466 bp ds-DNA     circular     14-NOV-2021
DEFINITION  .
KEYWORDS    "Antibiotic Resistance"                                            
FEATURES             Location/Qualifiers
     primer          236..260
                     /label="GA_seq fwd 1"
                     /note="sequence: GATGTGCTGCAAGGCGATTAAGTTG"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          270..292
                     /label="[Eurofins] M13 uni (-43)"
                     /note="sequence: AGGGTTTTCCCAGTCACGACGTT"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(290..309)
                     /label="GA MS2 rev"
                     /note="sequence: ctGGTCTCtCCTCACTGGCCGTCGTTTTACAAC"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer_bind     292..309
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          292..309
                     /label="[Eurofins] M13 uni (-21)"
                     /note="sequence: TGTAAAACGACGGCCAGT"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    310..313
                     /label="gg overlap"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          310..336
                     /label="pACYC_MS2_PLP FWD (2)"
                     /note="sequence: GTGGGTCTCCGAGGtaatacgactcactataggggaa"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer_bind     313..333
                     /label="T7"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     promoter        314..332
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     314..333
                     /label="T7"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    314..444
                     /label="T7 from pET28a"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_binding    333..355
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     protein_bind    333..357
                     /label="lac operator"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        366..395
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     RBS             415..437
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(417..448)
                     /label="pACYC_MS2_PLP REV (2)"
                     /note="sequence: GTGGGTCTCCGCCAatttaaatctccttcttaaagttaaaca"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          445..464
                     /label="pACYC_MS2_PLP FWD (3)"
                     /note="sequence: GTGGGTCTCGTGGCTATCGCTGTAGGTAGC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    445..465
                     /label="Primer MS2 NcoI"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    445..2512
                     /label="Insert"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             494..1675
                     /label="Maturase"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    529..529
                     /label="g->a (remove BsaI)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          1028..1051
                     /label="MS2_PLP seq 2"
                     /note="sequence: cttgccctaaacgaagatcgaaag"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1078..1078
                     /label="g->a (remove BspMI)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    1324..1324
                     /label="t->c (remove BsmBI)"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          1618..1641
                     /label="MS2_PLP seq 3"
                     /note="sequence: ctcgatggtccataccttagatgc"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    1699..2088
                     /label="Coat Protein"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    2034..2034
                     /label="c->t (remove BsaI)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    2083..2100
                     /label="Deletion Around Here to Restore Capsid Assembly"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    2092..2094
                     /label="Start of second coat dimer"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    2131..2148
                     /label="His Tag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    2442..2442
                     /label="c->t (remove BsaI)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    complement(2493..2512)
                     /label="Primer MS2 NotI"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(2493..2512)
                     /label="pACYC_MS2_PLP REV (3)"
                     /note="sequence: GTGGGTCTCGggTGGCCGGCGTCTATTAGTAG"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          2511..2544
                     /label="pACYC_MS2_PLP FWD (4)"
                     /note="sequence: CAccgctgagcaataactagcataaccccttggg"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer_bind     complement(2513..2531)
                     /label="T7 Term"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(2515..2548)
                     /label="pACYC_MS2_PLP REV (4)"
                     /note="sequence: aggccccaaggggttatgctagttattgctcagc"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     terminator      2527..2542
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     terminator      2543..2574
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          2545..2573
                     /label="pACYC_MS2_PLP FWD (5)"
                     /note="sequence: gcctctaaacgggtcttgaggggtttttt"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(2549..2577)
                     /label="pACYC_MS2_PLP REV (5)"
                     /note="sequence: TTAcaaaaaacccctcaagacccgtttag"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    2575..2577
                     /label="gg overlap"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    join(2578..4466,1..309)
                     /label="Backbone"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          2578..2595
                     /label="GA MS2 fwd"
                     /note="sequence: ctGGTCTCtGTAATTTCACTGCCCGCTTTCC"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     rep_origin      complement(2726..3393)
                     /label="Col\E1\origin"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     rep_origin      complement(2764..3446)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          complement(2769..2790)
                     /label="HIV Seq Fwd"
                     /note="sequence: ctatggaaaaacgccagcaacg"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          3397..3416
                     /label="MS2_PLP seq 5"
                     /note="sequence: GTCTGACGCTCAGTGGAACG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     CDS             complement(3541..4338)
                     /label="KanR"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
ORIGIN
        1 CTAAATTGTA AGCGTTAATA TTTTGTTAAA ATTCGCGTTA AATTTTTGTT AAATCAGCTC
       61 ATTTTTTAAC CAATAGGCCG AAATCGGCAA AATCCCTTAT AAATCAAAAG AATAGACCGA
      121 GATAGGGTTG AGTGGCCGCT ACAGGGCGCT CCCATTCGCC ATTCAGGCTG CGCAACTGTT
      181 GGGAAGGGCG TTTCGGTGCG GGCCTCTTCG CTATTACGCC AGCTGGCGAA AGGGGGATGT
      241 GCTGCAAGGC GATTAAGTTG GGTAACGCCA GGGTTTTCCC AGTCACGACG TTGTAAAACG
      301 ACGGCCAGTG AGGtaatacg actcactata ggggaattgt gagcggataa caattcccct
      361 ctagatttac actttatgct tccggctcgt atgtttctag agaattcaaa taattttgtt
      421 taactttaag aaggagattt aaatTGGCTA TCGCTGTAGG TAGCCGGAAT TCCATTCCTA
      481 GGAGGTTTGA CCTgtgcgag cttttagtac ccttgatagg gagaacgaaa ccttcgtccc
      541 ctccgttcgc gtttacgcgg acggtgagac tgaagataac tcattctctt taaaatatcg
      601 ttcgaactgg actcccggtc gttttaactc gactggggcc aaaacgaaac agtggcacta
      661 cccctctccg tattcacggg gggcgttaag tgtcacatcg atagatcaag gtgcctacaa
      721 gcgaagtggg tcatcgtggg gtcgcccgta cgaggagaaa gccggtttcg gcttctccct
      781 cgacgcacgc tcctgctaca gcctcttccc tgtaagccaa aacttgactt acatcgaagt
      841 gccgcagaac gttgcgaacc gggcgtcgac cgaagtcctg caaaaggtca cccagggtaa
      901 ttttaacctt ggtgttgctt tagcagaggc caggtcgaca gcctcacaac tcgcgacgca
      961 aaccattgcg ctcgtgaagg cgtacactgc cgctcgtcgc ggtaattggc gccaggcgct
     1021 ccgctacctt gccctaaacg aagatcgaaa gtttcgatca aaacacgtgg ccggcagatg
     1081 gttggagttg cagttcggtt ggttaccact aatgagtgat atccagggtg catatgagat
     1141 gcttacgaag gttcaccttc aagagtttct tcctatgaga gccgtacgtc aggtcggtac
     1201 taacatcaag ttagatggcc gtctgtcgta tccagctgca aacttccaga caacgtgcaa
     1261 catatcgcga cgtatcgtga tatggtttta cataaacgat gcacgtttgg catggttgtc
     1321 gtccctaggt atcttgaacc cactaggtat agtgtgggaa aaggtgcctt tctcattcgt
     1381 tgtcgactgg ctcctacctg taggtaacat gctcgagggc cttacggccc ccgtgggatg
     1441 ctcctacatg tcaggaacag ttactgacgt aataacgggt gagtccatca taagcgttga
     1501 cgctccctac gggtggactg tggagagaca gggcactgct aaggcccaaa tctcagccat
     1561 gcatcgaggg gtacaatccg tatggccaac aactggcgcg tacgtaaagt ctcctttctc
     1621 gatggtccat accttagatg cgttagcatt aatcaggcaa cggctctcta gatagAGCCC
     1681 TCAACCGGAG TTTGAAGCAT GGCTTCTAAC TTTACTCAGT TCGTTCTCGT CGACAATGGC
     1741 GGAACTGGCG ACGTGACTGT CGCCCCAAGC AACTTCGCTA ACGGGGTCGC TGAATGGATC
     1801 AGCTCTAACT CGCGTTCACA GGCTTACAAA GTAACCTGTA GCGTTCGTCA GAGCTCTGCG
     1861 CAGAATCGCA AATACACCAT CAAAGTCGAG GTGCCTAAAG TGGCAACCCA GACTGTTGGT
     1921 GGTGTAGAGC TTCCTGTAGC CGCATGGCGT TCGTACTTAA ATATGGAACT AACCATTCCA
     1981 ATTTTCGCTA CGAATTCCGA CTGCGAGCTT ATTGTTAAGG CAATGCAAGG TCTTCTAAAA
     2041 GATGGAAACC CGATTCCCTC AGCAATCGCA GCAAACTCCG GCATCTACGC TAACTTTACT
     2101 CAGTTCGTTC TCGTCGACAA TGGCGGTACC CATCACCATC ACCATCACGG TACCGGCGAC
     2161 GTGACTGTCG CCCCAAGCAA CTTCGCTAAC GGGGTCGCTG AATGGATCAG CTCTAACTCG
     2221 CGTTCACAGG CTTACAAAGT AACCTGTAGC GTtCGTCAGA GCTCTGCGCA GAATCGCAAA
     2281 TACACCATCA AAGTCGAGGT GCCTAAAGTG GCAACCCAGA CTGTTGGTGG TGTAGAGCTT
     2341 CCTGTAGCCG CATGGCGTTC GTACTTAAAT ATGGAACTAA CCATTCCAAT TTTCGCTACG
     2401 AATTCCGACT GCGAGCTTAT TGTTAAGGCA ATGCAAGGTC TTCTAAAAGA TGGAAACCCG
     2461 ATTCCCTCAG CAATCGCAGC AAACTCCGGC ATCTACTAAT AGACGCCGGC CAccgctgag
     2521 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgTAATTT
     2581 CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAACAT GGTCATAGCT
     2641 GTTTCCTTGC GTATTGGGCG CTCTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG
     2701 TTCGGGTAAA GCCTGGGGTG CCTAATGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA
     2761 AAAGGCCGCG TTGCTGGCGT TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA
     2821 TCGACGCTCA AGTCAGAGGT GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC
     2881 CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC
     2941 CGCCTTTCTC CCTTCGGGAA GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG
     3001 TTCGGTGTAG GTCGTTCGCT CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA
     3061 CCGCTGCGCC TTATCCGGTA ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC
     3121 GCCACTGGCA GCAGCCACTG GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC
     3181 AGAGTTCTTG AAGTGGTGGC CTAACTACGG CTACACTAGA AGAACAGTAT TTGGTATCTG
     3241 CGCTCTGCTG AAGCCAGTTA CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA
     3301 AACCACCGCT GGTAGCGGTG GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA
     3361 AGGATCTCAA GAAGATCCTT TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA
     3421 CTCACGTTAA GGGATTTTGG TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT
     3481 AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG
     3541 TTATTAGAAA AATTCATCCA GCAGACGATA AAACGCAATA CGCTGGCTAT CCGGTGCCGC
     3601 AATGCCATAC AGCACCAGAA AACGATCCGC CCATTCGCCG CCCAGTTCTT CCGCAATATC
     3661 ACGGGTGGCC AGCGCAATAT CCTGATAACG ATCCGCCACG CCCAGACGGC CGCAATCAAT
     3721 AAAGCCGCTA AAACGGCCAT TTTCCACCAT AATGTTCGGC AGGCACGCAT CACCATGGGT
     3781 CACCACCAGA TCTTCGCCAT CCGGCATGCT CGCTTTCAGA CGCGCAAACA GCTCTGCCGG
     3841 TGCCAGGCCC TGATGTTCTT CATCCAGATC ATCCTGATCC ACCAGGCCCG CTTCCATACG
     3901 GGTACGCGCA CGTTCAATAC GATGTTTCGC CTGATGATCA AACGGACAGG TCGCCGGGTC
     3961 CAGGGTATGC AGACGACGCA TGGCATCCGC CATAATGCTC ACTTTTTCTG CCGGCGCCAG
     4021 ATGGCTAGAC AGCAGATCCT GACCCGGCAC TTCGCCCAGC AGCAGCCAAT CACGGCCCGC
     4081 TTCGGTCACC ACATCCAGCA CCGCCGCACA CGGAACACCG GTGGTGGCCA GCCAGCTCAG
     4141 ACGCGCCGCT TCATCCTGCA GCTCGTTCAG CGCACCGCTC AGATCGGTTT TCACAAACAG
     4201 CACCGGACGA CCCTGCGCGC TCAGACGAAA CACCGCCGCA TCAGAGCAGC CAATGGTCTG
     4261 CTGCGCCCAA TCATAGCCAA ACAGACGTTC CACCCACGCT GCCGGGCTAC CCGCATGCAG
     4321 GCCATCCTGT TCAATCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA
     4381 TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC
     4441 GCGCACATTT CCCCGAAAAG TGCCAC
//