LOCUS pET-28a_(+)_RRF 5782 bp ds-DNA circular 23-MAR-2021 DEFINITION . FEATURES Location/Qualifiers primer 69..91 /label="pGEX_3_primer" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 feature complement(85..276) /label="ROP" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 gene complement(751..1026) /label="tet (576 - 851)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 regulatory complement(1085..2176) /label="lacI" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 gene complement(2259..2522) /label="tet (300 - 563)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 primer 2451..2470 /label="pBRrevBam_primer" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e promoter 2554..2572 /label="T7_promoter" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 regulatory 2572..2599 /label="lacO" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac feature 2617..2633 /label="T7_transl_en_RBS" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 misc_feature complement(2625..2643) /label="Tentative pET28 anneal FOR" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 2641..2643 /label="Start Codon" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 CDS 2641..3230 /label="Translation 2641-3230" gene 2644..3195 /label="frr gene" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 CDS 2644..3195 /label="frr CDS (ribosome recycling factor)" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 CDS 2644..3195 /label="Translation 2644-3195" misc_feature 3196..3214 /label="pET28 anneal REV" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 CDS 3196..3216 /label="Translation 3196-3216" terminator 3224..3352 /label="T7_terminator" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 3228..3230 /label="Stop Codon" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 primer complement(3266..3284) /label="T7_Terminal_primer" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 gene complement(3912..4727) /label="KanR2" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 origin 4833..5452 /label="pBR322_origin" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 ORIGIN 1 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 61 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 121 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc 181 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg 241 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt 301 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac 361 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat 421 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt 481 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca 541 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt 601 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg 661 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag 721 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc 781 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa 841 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat 901 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag 961 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct 1021 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt 1081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 1141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca 1201 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc 1261 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga 1321 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc 1381 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca 1441 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca 1501 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt 1561 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg 1621 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat 1681 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa 1741 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga 1801 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc 1861 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa 1921 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca 1981 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg 2041 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca 2101 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg 2161 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa 2221 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc 2281 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt 2341 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg 2401 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg 2461 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg 2521 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt 2581 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc 2641 atgattagcg atatcagaaa agatgctgaa gtacgcatgg acaaatgcgt agaagcgttc 2701 aaaacccaaa tcagcaaaat acgcacgggt cgtgcttctc ccagcctgct ggatggcatt 2761 gtcgtggaat attacggcac gccgacgccg ctgcgtcagc tggcaagcgt aacggtagaa 2821 gattcccgta cactgaaaat caacgtgttt gatcgttcaa tgtctccggc cgttgaaaaa 2881 gcgattatgg cgtccgatct tggcctgaac ccgaactctg cgggtagcga catccgtgtt 2941 ccgctgccgc cgctgacgga agaacgtcgt aaagatctga ccaaaatcgt tcgtggtgaa 3001 gcagaacaag cgcgtgttgc agtacgtaac gtgcgtcgtg acgcgaacga caaagtgaaa 3061 gcactgttga aagataaaga gatcagcgaa gacgacgatc gccgttctca ggacgatgta 3121 cagaaactga ctgatgctgc aatcaagaaa attgaagcgg cgctggcaga caaagaagca 3181 gaactgatgc agttccacca ccaccaccac cactgagatc cggctgctaa caaagcccga 3241 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc 3301 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat 3361 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 3421 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 3481 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 3541 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 3601 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3661 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 3721 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3781 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact tttcggggaa 3841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 3901 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca 3961 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg 4021 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca 4081 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca 4141 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt 4201 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc 4261 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa 4321 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct 4381 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt 4441 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc 4501 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca 4561 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct 4621 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa 4681 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta 4741 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt 4801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 4981 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 5461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 5521 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 5581 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct 5641 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc 5701 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 5761 ctgcgccccg acacccgcca ac //