LOCUS       PgyrA_RBS1_P48_His-cpsA 4204 bp ds-DNA     circular     30-NOV-2021
DEFINITION  .
COMMENT     Normal PCR (for backbone as well) sfGFP LK Bff-m:
            AGGACAACC*TGTATTTCC*agggG       100nm sfGFP LK Brr-m:
            GGAAATACA*GGTTGTCCT*tatA        100nm Primers including barcodes
            (for cpsADH) sfGFP LK cpsADH F:
            AGGACAACC*TGTATTTCC*agggGTCAATCCCGTCGTCCCA cpsADH N31 R:
            aaaccct*tcagccca*tcAcgggttaaagacgacgcg Barcoding (Future use) sfGFP
            LK RG-Boligo
            ccctGGAAATACAGGTTGTCCTagcttgAGGACAACCTGTATTTCCagggG     25nm sfGFP
            LK LA-Boligo      ataAGGACAACCTGTATTTCCagcttgGGAAATACAGGTTGTCCTtatA
            25nm F primer CATTCTGACCTGCACGTGAT R primer CACGTCATCCAGGTCGT
            Forward Primer with Barcode Plasmid library Q used in
            transformation ------                         F1 primer
            AGCTAGATGGCATTCTGACCTGCACGTGAT Library Q cells (liquid culture, 30
            C, 48 h) ---                        F2 primer
            TACCCGGGGCCATTCTGACCTGCACGTGAT Library Q+P48 cells (99/1 liquid
            culture, 30 C, 48h) -          F3 primer
            GAGCTAGTCGCATTCTGACCTGCACGTGAT Library Q+P48 cells (90/10 liquid
            culture, 30 C, 48h) -         F4 primer
            GACAATGCTACATTCTGACCTGCACGTGAT Library Q+P48 cells (50/50 liquid
            culture, 30 C, 48h) -         F5 primer
            CCCTAGACCGCATTCTGACCTGCACGTGAT P48 cells (liquid culture, 30 C,
            48h) -                                         F6 primer
            CGCTAGCTCACATTCTGACCTGCACGTGAT P48 plasmid (used in transformation)
            -                                  F7 primer
            TTCCTAGCGCCATTCTGACCTGCACGTGAT Reverse Primer with Barcode Plasmid
            library Q used in transformation ------                         R1
            primer       GGAACGGCTGTGCAGTCACACACGTCATCCAGGTCGT Library Q cells
            (liquid culture, 30 C, 48 h) ---                        R2 primer
            AGAAGATTGAGGTGTCCTAACACGTCATCCAGGTCGT Library Q+P48 cells (99/1
            liquid culture, 30 C, 48h) -          R3 primer
            ACTTACGCGGCCATAACACCCACGTCATCCAGGTCGT Library Q+P48 cells (90/10
            liquid culture, 30 C, 48h) -         R4 primer
            TTAGCCGTCTCGGGGGAATACACGTCATCCAGGTCGT Library Q+P48 cells (50/50
            liquid culture, 30 C, 48h) -         R5 primer
            AGTGACCTATGCACCAGACGCACGTCATCCAGGTCGT P48 cells (liquid culture, 30
            C, 48h) -                                         R6 primer
            CCGCTTGCGATTTCGACCAACACGTCATCCAGGTCGT P48 plasmid (used in
            transformation) -                                  R7 primer
            CGGGCCCCAACACGTCATCCAGGTCGT
FEATURES             Location/Qualifiers
     misc_feature    128..128
                     /label="Conserved T"
     misc_feature    128..154
                     /label="N21A"
     misc_feature    129..153
                     /label="N21A barcode design"
     misc_feature    154..154
                     /label="Conserved G"
     misc_feature    155..940
                     /label="aadA"
     CDS             155..940
                     /label="Translation 155-940"
                     /translation="REAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    941..941
                     /label="Conserved T"
     misc_feature    941..965
                     /label="N22A"
     misc_feature    942..964
                     /label="N22A barcode design"
     misc_feature    965..965
                     /label="Conserved G"
     misc_feature    1064..1084
                     /label="N22"
     CDS             1137..1725
                     /label="pMB1"
     misc_feature    1973..1993
                     /label="N21"
     misc_feature    1994..2193
                     /label="PgyrA"
     CDS             1994..2193
                     /label="Translation 1994-2193"
                     /translation="EATASNLAISSWSTLWFSGDFSAFIGTSTP*LARQTSISGIGCE*SV*VYLKLRGCVIICDL*IRD"
     misc_feature    2066..2075
                     /label="cspa trans start"
     misc_feature    2177..2183
                     /label="Ppnp trans start"
     misc_feature    2194..2236
                     /label="RBS1"
     CDS             2237..2254
                     /label="6xHis"
     CDS             2255..2962
                     /label="Translation 2255-2962"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAEFEIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDDHYLSTESVLSKDPNEDRDHMVLLEFVTAAGIDLGMDEL"
     misc_feature    2255..2968
                     /label="sfGFP(-15)_m"
     misc_feature    2963..2989
                     /label="Linker barcode"
     CDS             2966..2968
                     /label="Translation 2966-2968"
                     /translation="K"
     misc_feature    2967..2984
                     /label="SE"
     CDS             2969..2988
                     /label="Translation 2969-2988"
                     /translation="DNLYFQ"
     misc_feature    2969..2988
                     /label="TEV protease cleavage site"
     CDS             2987..2989
                     /label="Translation 2987-2989"
                     /translation="G"
     misc_feature    2990..3994
                     /label="cpsadh"
     CDS             2990..3994
                     /label="Translation 2990-3994"
                     /translation="SIPSSQYGFVFNKQSGLNLRNDLPVHKPKAGQLLLKVDAVGLCHSDLHVIYEGLDCGDNYVMGHEIAGTVAAVGDDVINYKVGDRVACVGPNGCGGCKYCRGAIDNVCKNAFGDWFGLGYDGGYQQYLLVTRPRNLSRIPDNVSADVAAASTDAVLTPYHAIKMAQVSPTSNILLIGAGGLGGNAIQVAKAFGAKVTVLDKKKEARDQAKKLGADAVYETLPESISPGSFSACFDFVSVQATFDVCQKYVEPKGVIMPVGLGAPNLSFNLGDLALREIRILGSFWGTTNDLDDVLKLVSEGKVKPVVRSAKLKELPEYIEKLRNNAYEGRVVFNP"
     misc_feature    3119..3138
                     /label="F Primer"
     misc_feature    3134..3148
                     /label="SE2"
     misc_feature    3149..3151
                     /label="L55NNK"
     misc_feature    3332..3334
                     /label="W116NNK"
     misc_feature    3824..3838
                     /label="SE1"
     misc_feature    3839..3844
                     /label="F285NNK W286NNK"
     misc_feature    complement(3855..3871)
                     /label="R Primer"
     misc_feature    3995..4015
                     /label="N31"
     misc_feature    4016..4171
                     /label="pheA terminator"
     misc_feature    4172..4190
                     /label="N23"
ORIGIN
        1 cttagatcta ttaccctgtt atccctactc gagttcatgt gcagctccat cagcaaaagg
       61 ggatgataag tttatcacca ccgactattt gcaacagtgc cgttgatcgt gctatgatcg
      121 actgatgtca tcagcggtgg agtgcaatgt catgagggaa gcggtgatcg ccgaagtatc
      181 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
      241 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
      301 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
      361 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
      421 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
      481 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
      541 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
      601 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
      661 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
      721 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
      781 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
      841 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
      901 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taagatgccg ctcgccagtc
      961 gattggctga gctcatgaag ttcctattcc gaagttccgc gaacgcgtaa aggatctagg
     1021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgTtcacac aacatagcca
     1081 cggGagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     1921 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatgctggat ccTTccctcg
     1981 actcacactt ggGgaggcga cggcttcaaa tttagcgatc tcttcgtggt ctacgttatg
     2041 gtttTccggc gatttttcgg cattcattgg cacttctact ccgtaattgg caagacaaac
     2101 gagtatatca ggcattggat gtgaataaag cgtataggtt tacctcaaac tgcgcggctg
     2161 tgttataatt tgcgaccttt gaatccggga tactagaaat aattttgttt aactttaaga
     2221 aggagatata catatgcacc atcatcatca tcatgtgagc aagggcgagg agctgttcac
     2281 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2341 gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac
     2401 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2461 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2521 cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg
     2581 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2641 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca acagccacaa
     2701 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccgaatttg aaattcgtca
     2761 taatgtggaa gatggcagcg tgcagctggc ggatcattat cagcagaata ccccgattgg
     2821 cgatggccca gtgctgctgc cggatgacca ctatctgagc accgaaagcg tgctgagcaa
     2881 agatccgaat gaagatcgtg atcatatggt cctgctggaa tttgtgaccg cggcaggcat
     2941 tgatctgggc atggatgaac tgtataaGga Caacctgtat ttCcagggGT CAATCCCGTC
     3001 GTCCCAATAC GGCTTTGTCT TCAATAAACA GTCTGGTCTG AATCTGCGTA ATGATCTGCC
     3061 GGTCCATAAA CCGAAAGCCG GCCAGCTGCT GCTGAAAGTC GATGCAGTGG GTCTGTGTCA
     3121 TTCTGACCTG CACGTGATTT ATGAAGGCCT GGATTGCGGT GACAACTACG TTATGGGCCA
     3181 TGAAATTGCA GGTACCGTTG CAGCAGTCGG TGATGACGTC ATCAACTATA AAGTGGGTGA
     3241 TCGTGTGGCG TGTGTTGGCC CGAATGGTTG CGGCGGTTGT AAATACTGCC GCGGCGCTAT
     3301 CGATAACGTG TGCAAAAATG CGTTTGGTGA TTGGTTCGGC CTGGGTTATG ACGGCGGTTA
     3361 TCAGCAATAC CTGCTGGTTA CCCGTCCGCG CAACCTGAGC CGTATTCCGG ATAATGTCTC
     3421 TGCTGACGTG GCAGCTGCGA GTACCGATGC GGTCCTGACG CCGTATCACG CCATCAAAAT
     3481 GGCACAGGTT TCACCGACCT CGAACATTCT GCTGATCGGT GCCGGCGGTC TGGGCGGTAA
     3541 TGCAATTCAA GTGGCCAAAG CATTTGGTGC GAAAGTTACG GTCCTGGATA AGAAAAAAGA
     3601 AGCGCGTGAT CAGGCGAAAA AACTGGGCGC TGATGCGGTT TATGAAACCC TGCCGGAAAG
     3661 CATTTCTCCG GGTAGTTTTT CCGCCTGTTT TGATTTCGTG TCAGTTCAGG CAACGTTCGA
     3721 CGTTTGCCAA AAATACGTCG AACCGAAAGG CGTTATCATG CCGGTCGGTC TGGGTGCTCC
     3781 GAACCTGTCG TTTAATCTGG GTGATCTGGC GCTGCGTGAA ATTCGCATCC TGGGCAGCTT
     3841 CTGGGGTACC ACGAACGACC TGGATGACGT GCTGAAACTG GTTTCCGAAG GCAAAGTGAA
     3901 ACCGGTGGTT CGTTCAGCCA AACTGAAAGA ACTGCCGGAA TATATCGAAA AACTGCGTAA
     3961 CAATGCATAC GAAGGTCGCG TCGTCTTTAA CCCGTgatgg gctgaagggt ttaaGCGAAG
     4021 ACGAACAATA AGGCCTCCCA AATCGGGGGG CCTTTTTTAT TGATAACAAA AAGGCAACAC
     4081 TATGACATCG GAAAACCCGT TACTGGCGCT GCGAGAGAAA ATCAGCGCGC TGGATGAAAA
     4141 ATTATTAGCA TTACTGGCAG AGCGGCGCGA Atcaaacaga aaggccatgg tcgacctgca
     4201 gaag
//