LOCUS       pET-28a_(+)_CK          6370 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     primer          2..24
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(18..209)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(684..959)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(1018..2109)
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     gene            complement(2192..2455)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          2384..2403
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        2487..2505
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      2505..2532
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         2550..2566
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(2558..2573)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             2574..3716
                     /label="Optimized ORF"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     CDS             2574..3751
                     /label="Translation 2574-3751"
     misc_feature    3717..3735
                     /label="pET28 anneal REV"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             3717..3737
                     /label="Translation 3717-3737"
     terminator      3745..3873
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    3749..3751
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(3787..3805)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(4433..5248)
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     origin          5354..5973
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
ORIGIN
        1 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
       61 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
      121 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
      181 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
      241 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
      301 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
      361 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
      421 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
      481 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
      541 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
      601 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
      661 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
      721 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
      781 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
      841 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
      901 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
      961 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     1021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1081 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     1141 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     1201 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     1261 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     1321 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     1381 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     1441 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     1501 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     1561 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     1621 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     1681 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     1741 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     1801 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     1861 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     1921 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     1981 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     2041 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     2101 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     2161 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     2221 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     2281 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     2341 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     2401 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     2461 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     2521 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accATGCCCT
     2581 TCGGAAATAC GCACAACAAG TACAAGCTGA ACTACAAGAG CGAGGAGGAG TACCCGGACC
     2641 TGAGCAAGCA CAACAACCAC ATGGCGAAAG TGTTAACTCC TGATCTGTAC AAGAAGCTGC
     2701 GCGACAAGGA GACCCCGAGC GGTTTTACCC TGGATGACGT TATTCAAACC GGCGTGGATA
     2761 ATCCTGGTCA CCCGTTCATC ATGACCGTGG GCTGTGTTGC AGGCGATGAA GAATCATACA
     2821 CCGTGTTCAA AGACCTGTTC GACCCGATCA TCCAGGACCG TCATGGCGGC TTCAAACCGA
     2881 CCGATAAGCA CAAAACCGAT CTGAACCACG AGAACCTGAA AGGCGGCGAT GATCTGGATC
     2941 CGCACTACGT TCTGAGCTCA AGAGTTCGTA CTGGTCGCAG CATTAAAGGC TACACTTTAC
     3001 CGCCGCACTG CTCACGTGGT GAAAGACGTG CAGTGGAAAA ACTGAGCGTT GAAGCGCTGA
     3061 ATTCATTAAC CGGCGAGTTC AAGGGCAAGT ACTACCCGCT GAAGAGCATG ACCGAACAGG
     3121 AGCAGCAGCA ATTGATCGAT GACCACTTCC TGTTCGACAA ACCGGTTTCA CCGCTTCTGT
     3181 TAGCGAGCGG TATGGCGAGA GATTGGCCTG ATGCACGTGG TATTTGGCAC AACGACAACA
     3241 AGAGCTTTCT GGTGTGGGTG AACGAGGAAG ACCATCTGCG CGTGATCAGC ATGGAGAAAG
     3301 GCGGCAACAT GAAAGAAGTG TTTCGCCGCT TTTGTGTGGG CCTGCAGAAG ATCGAGGAGA
     3361 TCTTCAAGAA GGCGGGTCAT CCGTTTATGT GGAACGAACA TCTGGGCTAC GTGCTGACTT
     3421 GTCCGAGCAA TTTAGGTACC GGCTTACGTG GTGGTGTGCA CGTTAAACTG GCGCATCTGA
     3481 GCAAACACCC GAAGTTCGAA GAAATCCTGA CCCGTTTACG CCTGCAGAAA CGTGGTACCG
     3541 GTGGCGTTGA TACTGCGGCA GTTGGTTCAG TTTTCGATAT CAGCAACGCG GATCGTTTAG
     3601 GTAGCAGCGA GGTGGAACAG GTGCAGCTGG TGGTTGATGG CGTGAAATTA ATGGTGGAGA
     3661 TGGAGAAGAA GCTGGAGAAG GGTCAGAGCA TCGACGACAT GATTCCGGCG CAGAAAcacc
     3721 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     3781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3841 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     3901 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3961 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4021 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4081 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4141 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4201 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4261 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4321 aatattaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4381 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     4441 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     4501 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     4561 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     4621 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     4681 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     4741 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     4801 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     4861 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     4921 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     4981 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     5041 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     5101 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     5161 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     5221 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     5281 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6121 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6181 atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     6241 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     6301 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6361 tgtgaccgtc
//