LOCUS pET-28a_(+)_RF1 6307 bp ds-DNA circular 23-MAR-2021 DEFINITION . FEATURES Location/Qualifiers gene complement(560..1375) /label="KanR2" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 origin 1481..2100 /label="pBR322_origin" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 primer 2499..2521 /label="pGEX_3_primer" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 feature complement(2515..2706) /label="ROP" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 gene complement(3181..3456) /label="tet (576 - 851)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 regulatory complement(3515..4606) /label="lacI" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 gene complement(4689..4952) /label="tet (300 - 563)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 primer 4881..4900 /label="pBRrevBam_primer" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e promoter 4984..5002 /label="T7_promoter" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 regulatory 5002..5029 /label="lacO" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac feature 5047..5063 /label="T7_transl_en_RBS" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 misc_feature complement(5055..5070) /label="Tentative pET28 anneal FOR" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 CDS 5071..6150 /label="prfA CDS (peptide chain release factor RF-1)" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 gene 5071..6150 /label="prfA gene" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 CDS 5071..6185 /label="Translation 5071-6185" misc_feature 6151..6169 /label="pET28 anneal REV" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 CDS 6151..6171 /label="Translation 6151-6171" terminator 6179..6307 /label="T7_terminator" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 6183..6185 /label="Stop Codon" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 primer complement(6221..6239) /label="T7_Terminal_primer" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc 3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa 3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc 3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac 3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca 3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta 3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca 3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa 3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt 3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg 3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca 3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta 3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg 4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat 4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct 4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg 4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat 4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc 4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca 4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg 4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt 4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg 4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct 4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga 4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg 4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc 4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg 4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg 4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga 4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa 5041 ttttgtttaa ctttaagaag gagatatacc atgaagcctt ctatcgttgc caaactggaa 5101 gccctgcatg aacgccatga agaagttcag gcgttgctgg gtgacgcgca aactatcgcc 5161 gaccaggaac gttttcgcgc attatcacgc gaatatgcgc agttaagtga tgtttcgcgc 5221 tgttttaccg actggcaaca ggttcaggaa gatatcgaaa ccgcacagat gatgctcgat 5281 gatcctgaaa tgcgtgagat ggcgcaggat gaactgcgcg aagctaaaga aaaaagcgag 5341 caactggaac agcaattaca ggttctgtta ctgccaaaag atcctgatga cgaacgtaac 5401 gccttcctcg aagtccgagc cggaaccggc ggcgacgaag cggcgctgtt cgcgggcgat 5461 ctgttccgta tgtacagccg ttatgccgaa gcccgccgct ggcgggtaga aatcatgagc 5521 gccagcgagg gtgaacatgg tggttataaa gagatcatcg ccaaaattag cggtgatggt 5581 gtgtatggtc gtctgaaatt tgaatccggc ggtcatcgcg tgcaacgtgt tcctgctacg 5641 gaatcgcagg gtcgtattca tacttctgct tgtaccgttg cggtaatgcc agaactgcct 5701 gacgcagaac tgccggacat caacccagca gatttacgca ttgatacttt ccgctcgtca 5761 ggggcgggtg gtcagcacgt taacaccacc gattcggcaa ttcgtattac tcacttgccg 5821 accgggattg ttgttgaatg tcaggacgaa cgttcacaac ataaaaacaa agctaaagca 5881 ctttctgttc tcggtgctcg catccacgct gctgaaatgg caaaacgcca acaggccgaa 5941 gcgtctaccc gtcgtaacct gctggggagt ggcgatcgca gcgaccgtaa ccgtacttac 6001 aacttcccgc aggggcgcgt taccgatcac cgcatcaacc tgacgctcta ccgcctggat 6061 gaagtgatgg aaggtaagct ggatatgctg attgaaccga ttatccagga acatcaggcc 6121 gaccaactgg cggcgttgtc cgagcaggaa caccaccacc accaccactg agatccggct 6181 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca 6241 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata 6301 tccggat //