LOCUS       pET-28a_(+)_HisRS       6499 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     origin          250..869
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          1268..1290
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(1284..1475)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(1950..2225)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(2284..3375)
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     gene            complement(3458..3721)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          3650..3669
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        3753..3771
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      3771..3798
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         3816..3832
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(3824..3842)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    3840..3842
                     /label="Start Codon"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             3840..5146
                     /label="Translation 3840-5146"
     gene            3843..5111
                     /label="hisS gene"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             3843..5111
                     /label="hisS CDS (histidyl tRNA synthetase)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     CDS             3843..5111
                     /label="Translation 3843-5111"
     misc_feature    5112..5130
                     /label="pET28 anneal REV"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             5112..5132
                     /label="Translation 5112-5132"
     terminator      5140..5268
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    5144..5146
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(5182..5200)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(join(5828..6499,1..144))
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
ORIGIN
        1 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
       61 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
      121 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
      181 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
      241 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      301 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      361 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
      421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      721 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      781 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      841 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      901 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      961 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     1021 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1081 cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     1141 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     1201 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1261 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     1321 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     1381 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     1441 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     1501 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     1561 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     1621 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     1681 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     1741 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     1801 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     1861 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     1921 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     1981 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     2041 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     2101 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     2161 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     2221 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     2281 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     2401 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     2461 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     2521 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     2581 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     2641 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     2701 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     2761 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     2821 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     2881 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     2941 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     3001 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     3061 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     3121 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     3181 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     3241 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     3301 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     3361 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     3421 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     3481 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     3541 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     3601 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     3661 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     3721 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     3781 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca
     3841 tggcaaaaaa cattcaagcc attcgcggca tgaacgatta cctgcctggc gaaacggcca
     3901 tctggcagcg cattgaaggc acactgaaaa acgtgctcgg cagctacggt tacagtgaaa
     3961 tccgcttgcc gattgtagag cagaccccgc tattcaaacg tgcgattggt gaagtcaccg
     4021 acgtggttga aaaagagatg tacacctttg aggatcgcaa tggcgacagc ctgactctgc
     4081 gccctgaagg gacggcgggc tgtgtacgcg ccggcatcga gcatggtctt ctgtacaatc
     4141 aggaacagcg tctgtggtat atcgggccga tgttccgtca cgagcgtccg cagaaagggc
     4201 gttatcgtca gttccatcag ttgggctgcg aagttttcgg tctgcaaggt ccggatatcg
     4261 acgctgaact gattatgctc actgcccgct ggtggcgcgc gctgggtatt tccgagcacg
     4321 taactcttga gctgaactct atcggttcgc tggaagcacg cgccaattac cgcgatgcgc
     4381 tggtggcatt ccttgagcag cataaagaaa agctggacga agactgcaaa cgccgcatgt
     4441 acactaaccc gctgcgcgtg ctggattcaa aaaatccgga agtgcaggcg cttctcaacg
     4501 acgctccggc attaggtgac tatctggacg aggaatctcg tgagcatttt gccggtctgt
     4561 gcaaactgct ggagagcgcg gggatcgctt acaccgtaaa ccagcgtctg gtgcgtggtc
     4621 tggattacta caaccgtacc gttttcgagt gggtgactaa cagtctcggc tcccagggca
     4681 ccgtgtgtgc aggcggtcgt tatgacggtc ttgtggaaca actgggcggt cgtgcaacac
     4741 cggctgtcgg ttttgctatg ggcctcgaac gtcttgtatt gttagtacag gccgttaatc
     4801 cggaatttaa agccgatcct gttgtcgata tatacctggt ggcttcaggt gctgatacac
     4861 aatctgcggc tatggcatta gctgagcgtc tgcgtgatga attaccgggc gtgaaattga
     4921 tgaccaacca cggcggcggc aactttaaga aacagtttgc ccgtgctgat aaatggggtg
     4981 cccgcgttgc tgtggtgctg ggtgagtctg aagtggctaa cggcacagca gtagtgaagg
     5041 atttgcgctc tggtgagcaa acggcagttg cgcaggatag cgtagccgcg catttgcgca
     5101 cgttactggg tcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     5161 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     5221 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     5281 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5341 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5401 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5461 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5521 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5581 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5641 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5701 cgcgaatttt aacaaaatat taacgtttac aatttcaggt ggcacttttc ggggaaatgt
     5761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     5821 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     5881 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     5941 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     6001 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     6061 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     6121 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     6181 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     6241 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     6301 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     6361 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     6421 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     6481 tcagaaacaa ctctggcgc
//