LOCUS pET-28a_(+)_ProRS 6943 bp ds-DNA circular 23-MAR-2021 DEFINITION . FEATURES Location/Qualifiers primer 56..78 /label="pGEX_3_primer" /ApEinfo_revcolor=#5b637e /ApEinfo_fwdcolor=#5b637e feature 72..263 /label="ROP" /ApEinfo_revcolor=#1ebdac /ApEinfo_fwdcolor=#1ebdac feature 738..1013 /label="tet(576-851)" /ApEinfo_revcolor=#ba9231 /ApEinfo_fwdcolor=#ba9231 gene 2246..2509 /label="tet (300-563)" /ApEinfo_revcolor=#ffe2b9 /ApEinfo_fwdcolor=#ffe2b9 primer 2438..2458 /label="pET28+ Reverse Primer" /ApEinfo_revcolor=#fffafa /ApEinfo_fwdcolor=#fffafa promoter 2541..2559 /label="T7 Promoter" /ApEinfo_revcolor=#c8498b /ApEinfo_fwdcolor=#c8498b regulatory 2559..2586 /label="lacI" /ApEinfo_revcolor=#c3dff3 /ApEinfo_fwdcolor=#c3dff3 regulatory 2559..2586 /label="lacO" /ApEinfo_revcolor=#fffbaa /ApEinfo_fwdcolor=#fffbaa feature 2604..2620 /label="T7 Ribosome Binding Site" /ApEinfo_revcolor=#e12222 /ApEinfo_fwdcolor=#e12222 gene 2628..4343 /label="Proline-tRNA ligase" /ApEinfo_revcolor=#0000ff /ApEinfo_fwdcolor=#0000ff primer 3438..3466 /label="seqFOR2 Proline-tRNA ligase C-tagged" /ApEinfo_revcolor=#f2c22e /ApEinfo_fwdcolor=#f2c22e primer 3951..3974 /label="seqFOR1 Proline-tRNA ligase C-tagged" /ApEinfo_revcolor=#f2c22e /ApEinfo_fwdcolor=#f2c22e feature 4344..4364 /label="His Tag and Stop Codon" /ApEinfo_revcolor=#ddb34e /ApEinfo_fwdcolor=#ddb34e terminator 4372..4500 /label="T7 Terminator" /ApEinfo_revcolor=#d24b19 /ApEinfo_fwdcolor=#d24b19 primer 4475..4499 /label="pET28+ Forward Primer" /ApEinfo_revcolor=#fffafa /ApEinfo_fwdcolor=#fffafa gene 5060..5875 /label="Kan R2" /ApEinfo_revcolor=#359797 /ApEinfo_fwdcolor=#359797 origin 5981..6600 /label="pBR322 Origin" /ApEinfo_revcolor=#9faa9a /ApEinfo_fwdcolor=#9faa9a ORIGIN 1 CCCTGACGGG CTTGTCTGCT CCCGGCATCC GCTTACAGAC AAGCTGTGAC CGTCTCCGGG 61 AGCTGCATGT GTCAGAGGTT TTCACCGTCA TCACCGAAAC GCGCGAGGCA GCTGCGGTAA 121 AGCTCATCAG CGTGGTCGTG AAGCGATTCA CAGATGTCTG CCTGTTCATC CGCGTCCAGC 181 TCGTTGAGTT TCTCCAGAAG CGTTAATGTC TGGCTTCTGA TAAAGCGGGC CATGTTAAGG 241 GCGGTTTTTT CCTGTTTGGT CACTGATGCC TCCGTGTAAG GGGGATTTCT GTTCATGGGG 301 GTAATGATAC CGATGAAACG AGAGAGGATG CTCACGATAC GGGTTACTGA TGATGAACAT 361 GCCCGGTTAC TGGAACGTTG TGAGGGTAAA CAACTGGCGG TATGGATGCG GCGGGACCAG 421 AGAAAAATCA CTCAGGGTCA ATGCCAGCGC TTCGTTAATA CAGATGTAGG TGTTCCACAG 481 GGTAGCCAGC AGCATCCTGC GATGCAGATC CGGAACATAA TGGTGCAGGG CGCTGACTTC 541 CGCGTTTCCA GACTTTACGA AACACGGAAA CCGAAGACCA TTCATGTTGT TGCTCAGGTC 601 GCAGACGTTT TGCAGCAGCA GTCGCTTCAC GTTCGCTCGC GTATCGGTGA TTCATTCTGC 661 TAACCAGTAA GGCAACCCCG CCAGCCTAGC CGGGTCCTCA ACGACAGGAG CACGATCATG 721 CGCACCCGTG GGGCCGCCAT GCCGGCGATA ATGGCCTGCT TCTCGCCGAA ACGTTTGGTG 781 GCGGGACCAG TGACGAAGGC TTGAGCGAGG GCGTGCAAGA TTCCGAATAC CGCAAGCGAC 841 AGGCCGATCA TCGTCGCGCT CCAGCGAAAG CGGTCCTCGC CGAAAATGAC CCAGAGCGCT 901 GCCGGCACCT GTCCTACGAG TTGCATGATA AAGAAGACAG TCATAAGTGC GGCGACGATA 961 GTCATGCCCC GCGCCCACCG GAAGGAGCTG ACTGGGTTGA AGGCTCTCAA GGGCATCGGT 1021 CGAGATCCCG GTGCCTAATG AGTGAGCTAA CTTACATTAA TTGCGTTGCG CTCACTGCCC 1081 GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA ACGCGCGGGG 1141 AGAGGCGGTT TGCGTATTGG GCGCCAGGGT GGTTTTTCTT TTCACCAGTG AGACGGGCAA 1201 CAGCTGATTG CCCTTCACCG CCTGGCCCTG AGAGAGTTGC AGCAAGCGGT CCACGCTGGT 1261 TTGCCCCAGC AGGCGAAAAT CCTGTTTGAT GGTGGTTAAC GGCGGGATAT AACATGAGCT 1321 GTCTTCGGTA TCGTCGTATC CCACTACCGA GATATCCGCA CCAACGCGCA GCCCGGACTC 1381 GGTAATGGCG CGCATTGCGC CCAGCGCCAT CTGATCGTTG GCAACCAGCA TCGCAGTGGG 1441 AACGATGCCC TCATTCAGCA TTTGCATGGT TTGTTGAAAA CCGGACATGG CACTCCAGTC 1501 GCCTTCCCGT TCCGCTATCG GCTGAATTTG ATTGCGAGTG AGATATTTAT GCCAGCCAGC 1561 CAGACGCAGA CGCGCCGAGA CAGAACTTAA TGGGCCCGCT AACAGCGCGA TTTGCTGGTG 1621 ACCCAATGCG ACCAGATGCT CCACGCCCAG TCGCGTACCG TCTTCATGGG AGAAAATAAT 1681 ACTGTTGATG GGTGTCTGGT CAGAGACATC AAGAAATAAC GCCGGAACAT TAGTGCAGGC 1741 AGCTTCCACA GCAATGGCAT CCTGGTCATC CAGCGGATAG TTAATGATCA GCCCACTGAC 1801 GCGTTGCGCG AGAAGATTGT GCACCGCCGC TTTACAGGCT TCGACGCCGC TTCGTTCTAC 1861 CATCGACACC ACCACGCTGG CACCCAGTTG ATCGGCGCGA GATTTAATCG CCGCGACAAT 1921 TTGCGACGGC GCGTGCAGGG CCAGACTGGA GGTGGCAACG CCAATCAGCA ACGACTGTTT 1981 GCCCGCCAGT TGTTGTGCCA CGCGGTTGGG AATGTAATTC AGCTCCGCCA TCGCCGCTTC 2041 CACTTTTTCC CGCGTTTTCG CAGAAACGTG GCTGGCCTGG TTCACCACGC GGGAAACGGT 2101 CTGATAAGAG ACACCGGCAT ACTCTGCGAC ATCGTATAAC GTTACTGGTT TCACATTCAC 2161 CACCCTGAAT TGACTCTCTT CCGGGCGCTA TCATGCCATA CCGCGAAAGG TTTTGCGCCA 2221 TTCGATGGTG TCCGGGATCT CGACGCTCTC CCTTATGCGA CTCCTGCATT AGGAAGCAGC 2281 CCAGTAGTAG GTTGAGGCCG TTGAGCACCG CCGCCGCAAG GAATGGTGCA TGCAAGGAGA 2341 TGGCGCCCAA CAGTCCCCCG GCCACGGGGC CTGCCACCAT ACCCACGCCG AAACAAGCGC 2401 TCATGAGCCC GAAGTGGCGA GCCCGATCTT CCCCATCGGT GATGTCGGCG ATATAGGCGC 2461 CAGCAACCGC ACCTGTGGCG CCGGTGATGC CGGCCACGAT GCGTCCGGCG TAGAGGATCG 2521 AGATCTCGAT CCCGCGAAAT TAATACGACT CACTATAGGG GAATTGTGAG CGGATAACAA 2581 TTCCCCTCTA GAAATAATTT TGTTTAACTT TAAGAAGGAG ATATACCATG CGTACTAGCC 2641 AATACCTGCT CTCCACTCTC AAGGAGACAC CTGCCGACGC CGAGGTGATC AGCCATCAGC 2701 TGATGCTGCG CGCCGGGATG ATCCGCAAGC TGGCCTCCGG GTTATATACC TGGCTGCCGA 2761 CCGGCGTGCG CGTTCTGAAA AAAGTCGAAA ACATCGTGCG TGAAGAGATG AACAACGCCG 2821 GTGCGATCGA GGTGTCGATG CCGGTGGTTC AGCCAGCCGA TTTGTGGCAA GAGAGTGGTC 2881 GTTGGGAACA GTACGGTCCG GAACTGCTGC GTTTTGTTGA CCGTGGCGAG CGTCCGTTCG 2941 TACTCGGCCC AACTCATGAA GAAGTTATCA CTGACCTGAT TCGTAACGAG CTTAGCTCTT 3001 ACAAACAGCT GCCGCTGAAC TTCTATCAGA TCCAGACCAA GTTCCGCGAC GAAGTGCGTC 3061 CGCGTTTCGG CGTCATGCGT TCCCGCGAAT TCCTGATGAA AGATGCTTAC TCTTTCCATA 3121 CTTCTCAGGA ATCCCTGCAG GAAACCTACG ATGCAATGTA TGCGGCCTAC AGCAAAATCT 3181 TCAGCCGCAT GGGGCTGGAT TTCCGCGCCG TACAAGCCGA CACCGGTTCT ATCGGCGGCA 3241 GCGCCTCTCA CGAATTCCAG GTGCTGGCGC AGAGCGGTGA AGACGATGTG GTCTTCTCCG 3301 ACACCTCTGA CTATGCAGCG AACATTGAAC TGGCAGAAGC TATCGCGCCG AAAGAACCGC 3361 GCGCTGCTGC TACCCAGGAA ATGACGCTGG TTGATACGCC GAACGCGAAA ACCATCGCGG 3421 AACTGGTTGA ACAGTTCAAT CTGCCGATTG AGAAAACGGT TAAGACTCTG CTGGTTAAAG 3481 CGGTTGAAGG CAGCAGCTTC CCGCAGGTTG CGCTGCTGGT GCGCGGTGAT CACGAGCTGA 3541 ACGAAGTTAA AGCAGAAAAA CTGCCGCAGG TTGCAAGCCC GCTGACTTTC GCGACCGAAG 3601 AAGAAATTCG TGCCGTGGTT AAAGCCGGTC CGGGTTCACT GGGTCCGGTA AACATGCCGA 3661 TTCCGGTGGT GATTGACCGT ACCGTTGCGG CGATGAGTGA TTTCGCTGCT GGTGCTAACA 3721 TCGATGGTAA ACACTACTTC GGCATCAACT GGGATCGCGA TGTCGCTACC CCGGAAGTTG 3781 CAGATATCCG TAACGTGGTG GCTGGCGATC CAAGCCCGGA TGGCCAGGGT AGGCTGCTGA 3841 TCAAACGTGG TATCGAAGTT GGTCACATCT TCCAGCTGGG TACCAAGTAC TCCGAAGCAC 3901 TGAAAGCCTC CGTACAGGGT GAAGATGGCC GTAACCAAAT CCTGACGATG GGTTGCTACG 3961 GTATCGGGGT AACGCGTGTG GTAGCTGCGG CGATTGAGCA GAACTACGAC GAACGAGGCA 4021 TCGTATGGCC TGACGCTATC GCGCCGTTCC AGGTGGCGAT TCTGCCGATG AACATGCACA 4081 AATCCTTCCG CGTACAAGAG CTTGCTGAGA AACTGTACAG CGAACTGCGT GCACAAGGTA 4141 TCGAAGTGCT GCTGGATGAC CGCAAAGAGC GTCCGGGCGT GATGTTTGCT GATATGGAAC 4201 TGATCGGTAT TCCGCACACT ATTGTGCTGG GCGACCGTAA CCTCGACAAC GACGATATCG 4261 AATATAAATA TCGTCGCAAC GGCGAGAAAC AGTTAATTAA GACTGGTGAC ATCGTCGAAT 4321 ATCTGGTGAA ACAGATTAAA GGCCACCACC ACCACCACCA CTGAGATCCG GCTGCTAACA 4381 AAGCCCGAAA GGAAGCTGAG TTGGCTGCTG CCACCGCTGA GCAATAACTA GCATAACCCC 4441 TTGGGGCCTC TAAACGGGTC TTGAGGGGTT TTTTGCTGAA AGGAGGAACT ATATCCGGAT 4501 TGGCGAATGG GACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG TGGTTACGCG 4561 CAGCGTGACC GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT TCTTCCCTTC 4621 CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGGC TCCCTTTAGG 4681 GTTCCGATTT AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG GTGATGGTTC 4741 ACGTAGTGGG CCATCGCCCT GATAGACGGT TTTTCGCCCT TTGACGTTGG AGTCCACGTT 4801 CTTTAATAGT GGACTCTTGT TCCAAACTGG AACAACACTC AACCCTATCT CGGTCTATTC 4861 TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA 4921 ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGTTT ACAATTTCAG GTGGCACTTT 4981 TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA 5041 TCCGCTCATG AATTAATTCT TAGAAAAACT CATCGAGCAT CAAATGAAAC TGCAATTTAT 5101 TCATATCAGG ATTATCAATA CCATATTTTT GAAAAAGCCG TTTCTGTAAT GAAGGAGAAA 5161 ACTCACCGAG GCAGTTCCAT AGGATGGCAA GATCCTGGTA TCGGTCTGCG ATTCCGACTC 5221 GTCCAACATC AATACAACCT ATTAATTTCC CCTCGTCAAA AATAAGGTTA TCAAGTGAGA 5281 AATCACCATG AGTGACGACT GAATCCGGTG AGAATGGCAA AAGTTTATGC ATTTCTTTCC 5341 AGACTTGTTC AACAGGCCAG CCATTACGCT CGTCATCAAA ATCACTCGCA TCAACCAAAC 5401 CGTTATTCAT TCGTGATTGC GCCTGAGCGA GACGAAATAC GCGATCGCTG TTAAAAGGAC 5461 AATTACAAAC AGGAATCGAA TGCAACCGGC GCAGGAACAC TGCCAGCGCA TCAACAATAT 5521 TTTCACCTGA ATCAGGATAT TCTTCTAATA CCTGGAATGC TGTTTTCCCG GGGATCGCAG 5581 TGGTGAGTAA CCATGCATCA TCAGGAGTAC GGATAAAATG CTTGATGGTC GGAAGAGGCA 5641 TAAATTCCGT CAGCCAGTTT AGTCTGACCA TCTCATCTGT AACATCATTG GCAACGCTAC 5701 CTTTGCCATG TTTCAGAAAC AACTCTGGCG CATCGGGCTT CCCATACAAT CGATAGATTG 5761 TCGCACCTGA TTGCCCGACA TTATCGCGAG CCCATTTATA CCCATATAAA TCAGCATCCA 5821 TGTTGGAATT TAATCGCGGC CTAGAGCAAG ACGTTTCCCG TTGAATATGG CTCATAACAC 5881 CCCTTGTATT ACTGTTTATG TAAGCAGACA GTTTTATTGT TCATGACCAA AATCCCTTAA 5941 CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA 6001 GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG 6061 GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC 6121 AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG 6181 AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC 6241 AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG 6301 CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC 6361 ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA 6421 AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT 6481 CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG 6541 CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG 6601 GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA 6661 TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC 6721 AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG CCTGATGCGG 6781 TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA TATATGGTGC ACTCTCAGTA 6841 CAATCTGCTC TGATGCCGCA TAGTTAAGCC AGTATACACT CCGCTATCGC TACGTGACTG 6901 GGTCATGGCT GCGCCCCGAC ACCCGCCAAC ACCCGCTGAC GCG //