LOCUS       pET-28a_(+)_T7RNAP      7918 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
KEYWORDS    "accession:addgene_vdb_2565"                                       
FEATURES             Location/Qualifiers
     origin          427..1046
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          1445..1467
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(1461..1652)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(2127..2402)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(2461..3552)
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     gene            complement(3635..3898)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          3827..3846
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        3930..3948
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      3948..3975
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         3993..4009
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(4001..4025)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    4017..4019
                     /label="Start Codon"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             4017..6742
                     /label="Translation 4017-6742"
     misc_feature    4029..4046
                     /label="His Tag"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    4056..4073
                     /label="Thrombin"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          4076..4076
                     /label="Xpress_fwd_primer"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(4076..4076)
                     /label="Tim Excises"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     gene            4077..6728
                     /label="T7 RNA Polymerase"
                     /ApEinfo_revcolor=#0000ff
                     /ApEinfo_fwdcolor=#0000ff
     primer          4769..4794
                     /label="seqFOR4 T7 RNA Polymerase N-tagged"
                     /ApEinfo_revcolor=#f2c22e
                     /ApEinfo_fwdcolor=#f2c22e
     primer          5284..5313
                     /label="seqFOR3 T7 RNA Polymerase N-tagged"
                     /ApEinfo_revcolor=#f2c22e
                     /ApEinfo_fwdcolor=#f2c22e
     primer          5789..5818
                     /label="seqFOR2 T7 RNA Polymerase N-tagged"
                     /ApEinfo_revcolor=#f2c22e
                     /ApEinfo_fwdcolor=#f2c22e
     primer          6319..6342
                     /label="seqFOR1 T7 RNA Polymerase N-tagged"
                     /ApEinfo_revcolor=#f2c22e
                     /ApEinfo_fwdcolor=#f2c22e
     terminator      6736..6864
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    6740..6742
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(6778..6796)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(join(7424..7918,1..321))
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
ORIGIN
        1 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
       61 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
      121 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
      181 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
      241 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
      301 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
      361 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
      421 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      481 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      541 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
      601 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      661 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      721 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      781 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      841 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      901 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      961 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1021 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1081 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1141 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1201 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1261 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     1321 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     1381 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1441 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     1501 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     1561 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     1621 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     1681 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     1741 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     1801 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     1861 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     1921 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     1981 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     2041 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     2101 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     2161 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     2221 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     2281 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     2341 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     2401 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     2461 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2521 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     2581 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     2641 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     2701 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     2761 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     2821 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     2881 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     2941 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     3001 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     3061 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     3121 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     3181 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     3241 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     3301 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     3361 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     3421 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     3481 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     3541 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     3601 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     3661 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     3721 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     3781 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     3841 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     3901 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     3961 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
     4021 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatATGA
     4081 ACACGATTAA CATCGCTAAG AACGACTTCT CTGACATCGA ACTGGCTGCT ATCCCGTTCA
     4141 ACACTCTGGC TGACCATTAC GGTGAGCGTT TAGCTCGCGA ACAGTTGGCC CTTGAGCATG
     4201 AGTCTTACGA GATGGGTGAA GCACGCTTCC GCAAGATGTT TGAGCGTCAA CTTAAAGCTG
     4261 GTGAGGTTGC GGATAACGCT GCCGCCAAGC CTCTCATCAC TACCCTACTC CCTAAGATGA
     4321 TTGCACGCAT CAACGACTGG TTTGAGGAAG TGAAAGCTAA GCGCGGCAAG CGCCCGACAG
     4381 CCTTCCAGTT CCTGCAAGAA ATCAAGCCGG AAGCCGTAGC GTACATCACC ATTAAGACCA
     4441 CTCTGGCTTG CCTAACCAGT GCTGACAATA CAACCGTTCA GGCTGTAGCA AGCGCAATCG
     4501 GTCGGGCCAT TGAGGACGAG GCTCGCTTCG GTCGTATCCG TGACCTTGAA GCTAAGCACT
     4561 TCAAGAAAAA CGTTGAGGAA CAACTCAACA AGCGCGTAGG GCACGTCTAC AAGAAAGCAT
     4621 TTATGCAAGT TGTCGAGGCT GACATGCTCT CTAAGGGTCT ACTCGGTGGC GAGGCGTGGT
     4681 CTTCGTGGCA TAAGGAAGAC TCTATTCATG TAGGAGTACG CTGCATCGAG ATGCTCATTG
     4741 AGTCAACCGG AATGGTTAGC TTACACCGCC AAAATGCTGG CGTAGTAGGT CAAGACTCTG
     4801 AGACTATCGA ACTCGCACCT GAATACGCTG AGGCTATCGC AACCCGTGCA GGTGCGCTGG
     4861 CTGGCATCTC TCCGATGTTC CAACCTTGCG TAGTTCCTCC TAAGCCGTGG ACTGGCATTA
     4921 CTGGTGGTGG CTATTGGGCT AACGGTCGTC GTCCTCTGGC GCTGGTGCGT ACTCACAGTA
     4981 AGAAAGCACT GATGCGCTAC GAAGACGTTT ACATGCCTGA GGTGTACAAA GCGATTAACA
     5041 TTGCGCAAAA CACCGCATGG AAAATCAACA AGAAAGTCCT AGCGGTCGCC AACGTAATCA
     5101 CCAAGTGGAA GCATTGTCCG GTCGAGGACA TCCCTGCGAT TGAGCGTGAA GAACTCCCGA
     5161 TGAAACCGGA AGACATCGAC ATGAATCCTG AGGCTCTCAC CGCGTGGAAA CGTGCTGCCG
     5221 CTGCTGTGTA CCGCAAGGAC AAGGCTCGCA AGTCTCGCCG TATCAGCCTT GAGTTCATGC
     5281 TTGAGCAAGC CAATAAGTTT GCTAACCATA AGGCCATCTG GTTCCCTTAC AACATGGACT
     5341 GGCGCGGTCG TGTTTACGCT GTGTCAATGT TCAACCCGCA AGGTAACGAT ATGACCAAAG
     5401 GACTGCTTAC GCTGGCGAAA GGTAAACCAA TCGGTAAGGA AGGTTACTAC TGGCTGAAAA
     5461 TCCACGGTGC AAACTGTGCG GGTGTCGATA AGGTTCCGTT CCCTGAGCGC ATCAAGTTCA
     5521 TTGAGGAAAA CCACGAGAAC ATCATGGCTT GCGCTAAGTC TCCACTGGAG AACACTTGGT
     5581 GGGCTGAGCA AGATTCTCCG TTCTGCTTCC TTGCGTTCTG CTTTGAGTAC GCTGGGGTAC
     5641 AGCACCACGG CCTGAGCTAT AACTGCTCCC TTCCGCTGGC GTTTGACGGG TCTTGCTCTG
     5701 GCATCCAGCA CTTCTCCGCG ATGCTCCGAG ATGAGGTAGG TGGTCGCGCG GTTAACTTGC
     5761 TTCCTAGTGA AACCGTTCAG GACATCTACG GGATTGTTGC TAAGAAAGTC AACGAGATTC
     5821 TACAAGCAGA CGCAATCAAT GGGACCGATA ACGAAGTAGT TACCGTGACC GATGAGAACA
     5881 CTGGTGAAAT CTCTGAGAAA GTCAAGCTGG GCACTAAGGC ACTGGCTGGT CAATGGCTGG
     5941 CTTACGGTGT TACTCGCAGT GTGACTAAGC GTTCAGTCAT GACGCTGGCT TACGGGTCCA
     6001 AAGAGTTCGG CTTCCGTCAA CAAGTGCTGG AAGATACCAT TCAGCCAGCT ATTGATTCCG
     6061 GCAAGGGTCT GATGTTCACT CAGCCGAATC AGGCTGCTGG ATACATGGCT AAGCTGATTT
     6121 GGGAATCTGT GAGCGTGACG GTGGTAGCTG CGGTTGAAGC AATGAACTGG CTTAAGTCTG
     6181 CTGCTAAGCT GCTGGCTGCT GAGGTCAAAG ATAAGAAGAC TGGAGAGATT CTTCGCAAGC
     6241 GTTGCGCTGT GCATTGGGTA ACTCCTGATG GTTTCCCTGT GTGGCAGGAA TACAAGAAGC
     6301 CTATTCAGAC GCGCTTGAAC CTGATGTTCC TCGGTCAGTT CCGCTTACAG CCTACCATTA
     6361 ACACCAACAA AGATAGCGAG ATTGATGCAC ACAAACAGGA GTCTGGTATC GCTCCTAACT
     6421 TTGTACACAG CCAAGACGGT AGCCACCTTC GTAAGACTGT AGTGTGGGCA CACGAGAAGT
     6481 ACGGAATCGA ATCTTTTGCA CTGATTCACG ACTCCTTCGG TACCATTCCG GCTGACGCTG
     6541 CGAACCTGTT CAAAGCAGTG CGCGAAACTA TGGTTGACAC ATATGAGTCT TGTGATGTAC
     6601 TGGCTGATTT CTACGACCAG TTCGCTGACC AGTTGCACGA GTCTCAATTG GACAAAATGC
     6661 CAGCACTTCC GGCTAAAGGT AACTTGAACC TCCGTGACAT CTTAGAGTCG GACTTCGCGT
     6721 TCGCGTAAga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     6781 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     6841 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     6901 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6961 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7021 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7081 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7141 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7201 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     7261 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     7321 gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7381 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     7441 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     7501 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     7561 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     7621 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     7681 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     7741 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     7801 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     7861 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctg
//