LOCUS       pET-28a_(+)_AlaRS       7897 bp ds-DNA     circular     23-MAR-2021
DEFINITION  .
KEYWORDS    "accession:addgene_vdb_2565"                                       
FEATURES             Location/Qualifiers
     origin          510..1129
                     /label="pBR322_origin"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          1528..1550
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     feature         complement(1544..1735)
                     /label="ROP"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     gene            complement(2210..2485)
                     /label="tet (576 - 851)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     regulatory      complement(2544..3635)
                     /label="lacI"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     gene            complement(3718..3981)
                     /label="tet (300 - 563)"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          3910..3929
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        4013..4031
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     regulatory      4031..4058
                     /label="lacO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     feature         4076..4092
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(4084..4108)
                     /label="Tentative pET28 anneal FOR"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    4100..4102
                     /label="Start Codon"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             4100..6804
                     /label="Translation 4100-6804"
     misc_feature    4112..4129
                     /label="His Tag"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    4139..4156
                     /label="Thrombin"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          4159..4159
                     /label="Xpress_fwd_primer"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(4159..4159)
                     /label="Tim Excises"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     gene            4160..6790
                     /label="alaS gene"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             4160..6790
                     /label="alaS CDS (alanyl-tRNA synthetase)"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     terminator      6798..6926
                     /label="T7_terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    6802..6804
                     /label="Stop Codon"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(6840..6858)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     gene            complement(join(7486..7897,1..404))
                     /label="KanR2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
ORIGIN
        1 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
       61 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
      121 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
      181 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
      241 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
      301 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
      361 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
      421 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
      481 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      541 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      601 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      661 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      721 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      781 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      841 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      901 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      961 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1021 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1081 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1141 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1201 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1261 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     1321 tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct
     1381 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     1441 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     1501 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     1561 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     1621 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     1681 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     1741 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     1801 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     1861 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     1921 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     1981 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     2041 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     2101 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     2161 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     2221 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     2281 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     2341 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     2401 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     2461 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     2521 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2581 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     2641 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     2701 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     2761 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     2821 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     2881 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     2941 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     3001 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     3061 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     3121 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     3181 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     3241 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     3301 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     3361 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     3421 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     3481 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     3541 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     3601 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     3661 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     3721 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     3781 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     3841 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     3901 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     3961 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
     4021 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
     4081 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
     4141 ggtgccgcgc ggcagccata tgagcaagag caccgctgag atccgtcagg cgtttctcga
     4201 ctttttccat agtaagggac atcaggtagt tgccagcagc tccctggtac cccataacga
     4261 cccaactttg ttgtttacca acgccgggat gaaccagttc aaggatgtgt tccttgggct
     4321 cgacaagcgt aattattccc gcgctaccac ttcccaacgc tgcgtgcgtg cgggtggtaa
     4381 acacaacgac ctggaaaacg tcggttacac cgcgcgtcac cataccttct tcgaaatgct
     4441 gggcaacttc agcttcggcg actatttcaa acacgatgcc attcagtttg catgggaact
     4501 gctgaccagc gaaaaatggt ttgccctgcc gaaagagcgt ctgtgggtta ccgtctatga
     4561 aagcgacgac gaagcctacg aaatctggga aaaagaagta gggatcccgc gcgaacgtat
     4621 tattcgcatc ggcgataaca aaggtgcgcc atacgcatct gacaacttct ggcagatggg
     4681 tgacactggt ccgtgcggcc cgtgcaccga aatcttctac gatcacggcg accacatttg
     4741 ggggggccct ccgggaagcc cggaagaaga cggcgaccgc tacattgaga tctggaacat
     4801 cgtcttcatg cagttcaacc gccaggccga tggcacgatg gaaccgctgc cgaagccgtc
     4861 tgtagatacc ggtatgggtc tggagcgtat tgctgcggtg ctgcaacacg ttaactctaa
     4921 ctatgacatc gacctgttcc gcacgctgat ccaggcggta gcgaaagtca ctggcgcaac
     4981 cgatctgagc aataaatcgc tgcgcgtaat cgctgaccac attcgttctt gtgcgttcct
     5041 gatcgcggat ggcgtaatgc cgtccaatga aaaccgtggt tatgtactgc gtcgtatcat
     5101 tcgtcgcgca gtgcgtcacg gtaatatgct cggcgcgaaa gaaaccttct tctacaaact
     5161 ggttggtccg ctgatcgacg ttatgggctc tgcgggtgaa gacctgaaac gccagcaggc
     5221 gcaggttgag caggtgctga agactgaaga agagcagttt gctcgtactc tggagcgcgg
     5281 tctggcgttg ctggatgaag agctggcaaa actttctggt gatacgctgg atggtgaaac
     5341 tgctttccgt ctgtacgaca cctatggctt cccggttgac ctgacggctg atgtttgtcg
     5401 tgagcgcaac atcaaagttg acgaagctgg ttttgaagct gcaatggaag agcagcgtcg
     5461 tcgcgcgcgc gaagccagcg gctttggtgc cgattacaac gcaatgatcc gtgttgacag
     5521 tgcatctgaa tttaaaggct atgaccatct ggaactgaac ggcaaagtga ctgcgctgtt
     5581 tgttgatggt aaagcggttg atgccatcaa tgcaggccag gaagctgtgg tcgtgctgga
     5641 tcaaacgcca ttctatgcgg aatccggcgg tcaggttggc gataaaggcg aactgaaagg
     5701 cgctaacttc tcctttgcgg tggaagatac gcagaaatac ggccaggcga ttggtcacat
     5761 cggtaaactt gctgcgggtt ctctgaaagt gggcgacgcg gtgcaggctg atgttgatga
     5821 ggctcgtcgc gcccgtattc gtctgaatca ctccgcaacg cacctgatgc acgctgcgct
     5881 gcgccaggtt ctgggtactc atgtatcgca gaaaggttca ctggttaacg acaaggtgct
     5941 gcgcttcgac ttctcacaca acgaagcgat gaaaccagaa gagattcgtg cggtcgaaga
     6001 cctggtgaac acacagattc gtcgcaattt gccgatcgaa accaacatca tggatctcga
     6061 agcggcgaaa gcgaaaggtg cgatggcgct gttcggcgag aagtatgatg agcgcgtacg
     6121 cgtgctgagc atgggcgatt tctctaccga gttgtgtggc ggtactcacg ccagccgcac
     6181 tggtgatatt ggtctgttcc gcatcatctc tgaatcgggt actgctgcag gcgttcgtcg
     6241 tatcgaagcg gtaaccggag aaggtgctat cgccaccgtt catgcagaca gcgatcgctt
     6301 aagcgaagtc gcgcatctgc tgaaaggcga tagcaataat ctggctgata aagtgcgctc
     6361 agtactggaa cgtacgcgtc agctggaaaa agagttacaa cagcttaaag aacaagctgc
     6421 cgcacaggag agcgcaaatc tttccagtaa ggcaattgat gttaatggtg ttaagctgtt
     6481 ggttagcgag cttagcggtg ttgagccgaa aatgttgcgt accatggttg acgatttaaa
     6541 aaatcagctg gggtcgacaa ttatcgtgct ggcaacggta gtcgaaggta aggtttctct
     6601 gattgcaggc gtatctaagg acgtcacaga tcgtgtgaaa gcaggggaac tgattggtat
     6661 ggtcgctcag caggtgggcg gcaagggtgg tggacgtcct gacatggcgc aagccggtgg
     6721 tacggatgct gcggccttac ctgcagcgtt agccagtgtg aaaggctggg tcagcgcgaa
     6781 attgcaataa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     6841 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     6901 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     6961 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7021 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7081 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7141 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7201 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7261 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7321 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7381 acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     7441 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     7501 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     7561 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     7621 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     7681 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     7741 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     7801 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     7861 aaatacgcga tcgctgttaa aaggacaatt acaaaca
//