LOCUS       pNB-Rep                 3604 bp ds-DNA     circular     15-APR-2025
DEFINITION  .
COMMENT     Imported using the Genbank importer. File name: 2 pCDF-MCS.gbk.txt
FEATURES             Location/Qualifiers
     misc_feature    184..189
                     /label="EcoRI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    190..219
                     /label="Truncated 2017mar28_200bpSpacer1"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    220..225
                     /label="HindIII"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    226..240
                     /label="Prefix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    241..275
                     /label="P(BBa_J23100)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    276..290
                     /label="Suffix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    291..320
                     /label="VL2017mar22_24"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    321..335
                     /label="Prefix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    complement(336..338)
                     /label="PAM"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(336..358)
                     /label="bssgRNA-1 _ ANTISENSE is identical (orthogonal set from Didovyk et al 2015)"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(339..358)
                     /label="orthogonal bssgRNA-1 _ ANTISENSE is identical (Didovyk et al 2015)"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    359..373
                     /label="Suffix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    374..403
                     /label="VL2017mar22_22"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    404..418
                     /label="Prefix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    419..438
                     /label="Csy4 cut site - Tsai et al 2014"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    424..438
                     /label="Csy4 minimal cut motif"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    438..438
                     /label="Csy4 cuts after this G"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    439..472
                     /label="RBS"
                     /ApEinfo_revcolor="#7ea7d8"
                     /ApEinfo_fwdcolor="#7ea7d8"
     misc_feature    460..465
                     /label="Shine-Dalgarno ?"
                     /ApEinfo_revcolor="#fdc68a"
                     /ApEinfo_fwdcolor="#fdc68a"
     gene            473..1186
                     /label="sfGFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             473..1252
                     /label="Translation 473-1252"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYKMSRRNTDAITIHSILDWIED**"
     misc_feature    1187..1252
                     /label="MarAn20 degradation tag (C-term)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    1253..1267
                     /label="Suffix"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1268..1273
                     /label="BamHI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1274..1303
                     /label="VL2017mar22_10"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1304..1311
                     /label="SbfI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1312..1372
                     /label="L3S2P21 Terminator"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1373..1572
                     /label="2018mar28_200bpSpacer2.5* (A->T change to remove AGCAGY)"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    1434..1434
                     /label="A->T change to remove AGCAGY"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1573..1578
                     /label="SalI"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     gene            complement(1994..2782)
                     /label="SpecR"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    complement(2922..3604)
                     /label="cloned fragment (expanded ori ???)"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     rep_origin      complement(2940..3528)
                     /label="pUC ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
ORIGIN
        1 ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
       61 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
      121 aggagctgac tgggttgCTC CTAGGATGCA GTGACTCGca catttccccg aaaagtgcca
      181 cctGAATTCG AAATCCCTGA AACTGAGACT GTAGAAAATA AGCTTCAGCC TGCGGTCCGG
      241 ttgacggcta gctcagtcct aggtacagtg ctagcTCGCT GGGACGCCCG AGATAGCCGT
      301 TACACAGGTG ACACTTATTT CAGCCTGCGG TCCGGccaCA CTAGAGTCTA GCTTGAGATC
      361 GCTGGGACGC CCGTAGAACT CTGTGAGGAT AAAGTCTCCC TTACAGCCTG CGGTCCGGGT
      421 TCACTGCCGT ATAGGCAGTA ATTTTGTTTA ACTTTAAGAA GGAGATATAC ATATGCGTAA
      481 AGGCGAAGAA CTGTTTACCG GTGTGGTTCC GATTCTGGTG GAACTGGACG GCGATGTTAA
      541 TGGTCATAAA TTCAGTGTTC GCGGCGAAGG TGAAGGCGAT GCGACGAACG GCAAACTGAC
      601 CCTGAAATTT ATCTGCACCA CGGGTAAACT GCCGGTCCCG TGGCCGACGC TGGTGACCAC
      661 GCTGACCTAT GGCGTTCAAT GTTTTGCGCG TTACCCGGAT CACATGAAAC AGCACGACTT
      721 TTTCAAATCG GCCATGCCGG AAGGCTATGT GCAGGAACGT ACGATTAGCT TTAAAGACGA
      781 TGGTACGTAT AAAACCCGCG CGGAAGTGAA ATTCGAAGGC GATACCCTGG TTAACCGTAT
      841 CGAACTGAAA GGTATCGATT TCAAAGAAGA CGGCAATATT CTGGGTCATA AACTGGAATA
      901 TAACTTCAAT TCCCACAACG TGTACATCAC CGCGGATAAA CAGAAAAACG GCATTAAAGC
      961 CAATTTCAAA ATCCGCCATA ATGTGGAAGA TGGTAGCGTT CAGCTGGCCG ACCACTATCA
     1021 GCAAAACACG CCGATTGGTG ATGGCCCGGT CCTGCTGCCG GACAATCACT ACCTGAGTAC
     1081 CCAGTCCGTG CTGTCAAAAG ATCCGAACGA AAAACGTGAC CACATGGTCC TGCTGGAATT
     1141 TGTGACGGCT GCGGGTATCA CCCACGGCAT GGACGAACTG TATAAAATGT CCCGCCGTAA
     1201 TACTGACGCC ATCACAATCC ACAGCATCCT GGATTGGATT GAAGACTAAT AATCGCTGGG
     1261 ACGCCCGGGA TCCAAGAGAT TTCTACACGA TTGAGCACTG TCTCCTGCAG GCTCGGTACC
     1321 AAATTCCAGA AAAGAGGCCT CCCGAAAGGG GGGCCTTTTT TCGTTTTGGT CCTAATAGAT
     1381 AAAGGATAGG TCTGGTAGTG TTGTTCGTTC TCGCAGGTAA ATCAATAATA CTCtGCAGTT
     1441 CCGTAGACTT TTCAGTGGGA CAGGGTAGCG ATAACAGATA GATTGTAATA AGACACAGTA
     1501 GGTGCTCGTA GTTGCGTGAA GAGAACCGCT CAGGAAATCC AGTCAGAAGT ATTGGTAATC
     1561 GTTGAAAACT CAGTCGACTG CTGCCACCGC TGAGCAATAA CTAGCATAAC CCCTTGGGGC
     1621 CTCTAAACGG GTCTTGAGGG GTTTTTTGCC AGGCATCAAA TAAAACGAAA GGCTCAGTCG
     1681 GAAGACTGGG CCTTTCGTTT TATCTGTTGT TTGTCGGTGA ACGCTCTCCT GAGTAGGACA
     1741 AATCCGCCGG GAGCGGATTT GAACGTTGTG AAGCAACGGC CCGGAGGGTG GCGGGCAGGA
     1801 CGCCCGCCAT AAACTGCCAG GCATCAAACT AAGCAGAAGG CCATCCTGAC GGATGGCCTT
     1861 TTTGCGTTTC AGATCTaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     1921 gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa
     1981 ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
     2041 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
     2101 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
     2161 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
     2221 gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt
     2281 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     2341 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
     2401 cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt
     2461 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     2521 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     2581 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     2641 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
     2701 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     2761 cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg
     2821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta
     2881 gctcactcgg tcgctacgct ccgggcgtga gactgcggcg gGGCCGCGTT GCTGGCGTTT
     2941 TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG
     3001 CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC
     3061 TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC
     3121 GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC
     3181 AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC
     3241 TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT
     3301 AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT
     3361 AACTACGGCT ACACTAGAAG AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC
     3421 TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT
     3481 TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG
     3541 ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC
     3601 ATGA
//