LOCUS       pET-28a_(+)_MK          5893 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     origin          139..758
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          1157..1179
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(1173..1364)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(1839..2114)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(2173..3264)
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     gene            complement(3347..3610)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          3539..3558
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        3642..3660
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      3660..3687
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         3705..3721
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(3713..3728)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             3729..4394
                     /label="Optimized ORF"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             3729..4429
                     /label="Translation 3729-4429"
     misc_feature    4395..4413
                     /label="pET28 anneal REV"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             4395..4415
                     /label="Translation 4395-4415"
     terminator      4423..4551
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    4427..4429
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(4465..4483)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(join(5111..5893,1..33))
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
ORIGIN
        1 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
       61 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      181 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      301 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      541 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      781 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      841 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      901 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      961 gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata
     1021 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     1081 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     1141 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     1201 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     1261 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     1321 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     1381 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     1441 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     1501 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     1561 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     1621 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     1681 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     1741 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     1801 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     1861 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     1921 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     1981 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     2041 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     2101 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     2161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     2281 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     2341 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     2401 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     2461 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     2521 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     2581 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     2641 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     2701 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     2761 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     2821 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     2881 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     2941 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     3001 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     3061 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     3121 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     3181 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     3241 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     3301 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     3361 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     3421 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     3481 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     3541 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     3601 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
     3661 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
     3721 gatataccAT GAGCAGCAGT GAGTCGATCC GTATGGTGCT GATTGGTCCT CCTGGTGCAG
     3781 GTAAAGGTAC TCAAGCACCG AATCTGCAGG AACGCTTTCA TGCGGCACAT TTAGCAACCG
     3841 GCGACATGTT ACGTAGCCAG ATCGCGAAAG GTACCCAGTT AGGTTTAGAG GCGAAGAAGA
     3901 TCATGGACCA GGGTGGTCTG GTGAGCGACG ACATTATGGT GAACATGATC AAGGACGAGC
     3961 TGACCAACAA TCCGGCGTGC AAAAACGGCT TCATCCTGGA CGGTTTTCCG CGTACTATTC
     4021 CTCAGGCGGA GAAATTAGAC CAGATGCTGA AGGAGCAGGG TACCCCTCTG GAAAAGGCGA
     4081 TCGAACTGAA AGTGGACGAC GAATTACTGG TTGCGCGTAT TACCGGTCGC CTGATTCATC
     4141 CGGCATCAGG TCGCTCATAC CACAAGATCT TTAATCCGCC GAAGGAAGAC ATGAAGGACG
     4201 ACGTTACCGG CGAAGCGCTG GTTCAGCGTA GCGATGATAA TGCAGATGCG CTGAAAAAAC
     4261 GTTTAGCGGC GTATCACGCA CAGACCGAGC CGATCGTGGA CTTCTACAAG AAAACCGGCA
     4321 TTTGGGCAGG TGTTGATGCA TCACAACCTC CGGCAACCGT TTGGGCGGAT ATCCTGAATA
     4381 AACTGGGCAA GGACcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     4441 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4501 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     4561 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4621 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4681 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4741 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4801 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4861 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4921 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4981 taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa
     5041 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5101 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     5161 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     5221 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     5281 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     5341 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     5401 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     5461 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     5521 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     5581 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     5641 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     5701 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     5761 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     5821 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     5881 ttaatcgcgg cct
//