LOCUS       pET-28a_(+)_RF3         6856 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
KEYWORDS    "accession:addgene_vdb_2565"                                       
FEATURES             Location/Qualifiers
     gene            complement(717..980)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          909..928
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        1012..1030
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      1030..1057
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         1075..1091
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(1083..1107)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    1099..1101
                     /label="Start Codon"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             1099..2762
                     /label="Translation 1099-2762"
     misc_feature    1111..1128
                     /label="His Tag"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    1138..1155
                     /label="Thrombin"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          1158..1158
                     /label="Xpress_fwd_primer"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(1158..1158)
                     /label="Tim Excises"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     gene            1159..2748
                     /label="prfC gene"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             1159..2748
                     /label="prfC CDS (peptide chain release factor RF-3)"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             1159..2748
                     /label="Translation 1159-2748"
     CDS             1159..2748
                     /label="Translation 1159-2748"
     CDS             1159..2748
                     /label="Translation 1159-2748"
     CDS             1159..2748
                     /label="Translation 1159-2748"
     CDS             1159..2748
                     /label="Translation 1159-2748"
     terminator      2756..2884
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    2760..2762
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(2798..2816)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(3444..4259)
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     origin          4365..4984
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          5383..5405
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(5399..5590)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(6065..6340)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(join(6399..6856,1..634))
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
ORIGIN
        1 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
       61 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
      121 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
      181 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
      241 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
      301 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
      361 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
      421 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
      481 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
      541 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
      601 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
      661 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
      721 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
      781 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
      841 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
      901 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
      961 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
     1021 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
     1081 ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg
     1141 gtgccgcgcg gcagccatat gacgttgtct ccttatttgc aagaggtggc gaagcgccgc
     1201 acttttgcca ttatttctca cccggacgcc ggtaagacta ccatcaccga gaaggtgctg
     1261 ctgttcggac aggccattca gaccgccggt acagtaaaag gccgtggttc caaccagcac
     1321 gctaagtcgg actggatgga gatggaaaag cagcgtggga tctccattac tacgtctgtg
     1381 atgcagtttc cgtatcacga ttgcctggtt aacctgctcg acaccccggg gcacgaagac
     1441 ttctcggaag atacctatcg taccctgacg gcggtggact gctgcctgat ggttatcgac
     1501 gccgcaaaag gtgttgaaga tcgtacccgt aagctgatgg aagttacccg tctgcgcgac
     1561 acgccgatcc tcacctttat gaacaaactt gaccgtgata tccgcgaccc gatggagctg
     1621 ctcgatgaag ttgagaacga gctgaaaatc ggctgtgcgc cgatcacctg gccgattggc
     1681 tgcggcaagc tgtttaaagg cgtttaccac ctttataaag acgaaaccta tctctatcag
     1741 agcggtaaag gccacaccat tcaggaagtc cgcattgtta aagggctgaa taacccggat
     1801 ctcgatgctg cggttggtga agatctggca cagcagctgc gtgacgaact ggaactggtg
     1861 aaaggcgcgt ctaacgagtt cgacaaagag ctgttccttg cgggcgaaat cactccggta
     1921 ttcttcggta ctgcgctggg taacttcggc gtcgatcata tgttggatgg cctggtggag
     1981 tgggcacctg cgccgatgcc gcgtcagact gatacccgta ccgtagaagc gagcgaagat
     2041 aaatttaccg gcttcgtatt taaaattcag gccaacatgg acccgaaaca ccgcgaccgc
     2101 gtggcgttta tgcgtgtggt gtccggtaaa tatgaaaaag gcatgaaact gcgccaggtg
     2161 cgcactgcga aagatgtggt gatctccgac gcgctgacct ttatggcggg tgaccgttcg
     2221 cacgttgaag aagcgtatcc gggcgatatc ctcggcctgc acaaccacgg caccattcag
     2281 atcggcgaca cctttaccca gggtgagatg atgaagttca ccggtattcc gaacttcgca
     2341 ccagaactgt tccgtcgtat ccgcctgaaa gatccgctga agcaaaaaca gctgctcaaa
     2401 gggctggtac agctttccga agagggcgcg gtgcaggtgt tccgtccaat ctccaacaac
     2461 gatctgatcg ttggtgcagt tggtgtgctg cagtttgatg tggtggtagc gcgcctgaag
     2521 agcgaataca acgttgaagc agtgtatgag tcagtcaacg ttgccactgc ccgctgggta
     2581 gaatgtgcag acgcgaagaa attcgaagag ttcaagcgta agaacgaaag ccaactggcg
     2641 cttgatggcg gcgataacct cgcttacatc gctaccagca tggtcaacct gcgcctggca
     2701 caggaacgtt atccggacgt tcagttccac cagacccgcg agcattaaga tccggctgct
     2761 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2821 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2881 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2941 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3001 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3061 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3121 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3181 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3241 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3301 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca
     3361 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3421 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
     3481 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     3541 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     3601 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     3661 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     3721 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     3781 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     3841 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     3901 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     3961 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     4021 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     4081 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     4141 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     4201 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
     4261 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     4321 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4441 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4501 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4561 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4621 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4741 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4801 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4861 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4921 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4981 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5041 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5101 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     5161 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc
     5221 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     5281 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5341 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5401 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     5461 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     5521 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     5581 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     5641 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     5701 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     5761 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     5821 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     5881 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     5941 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     6001 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     6061 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     6121 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     6181 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     6241 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     6301 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     6361 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6481 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     6541 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     6601 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     6661 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     6721 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     6781 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     6841 gctatcggct gaattt
//