LOCUS       Exported                6354 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses E. coli HPII catalase with TAG stop codon at site 415 and 
            N-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-HPII TAG415
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  TITLE     Bacterial expression plasmids for genetic code expansion laboratory 
            course
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6354)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6354
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        11..295
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     218..237
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             341..358
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             362..394
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             398..421
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2765..2782)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2935..3021
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3113..3140
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3159..3250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3251..4111
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3469..3488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4282..4870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4771..4790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5024..5041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5056..5196
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5122..5141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5282..5304
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5460..6338)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
       61 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
      121 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
      181 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
      241 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
      301 ttttttgggc taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg
      361 tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa
      421 ggatcgatgg ggatccgagc tcgagtcgca acataacgaa aagaacccac atcagcacca
      481 gtcaccacta cacgattcca gcgaagcgaa accggggatg gactcactgg cacctgagga
      541 cggctctcat cgtccagcgg ctgaaccaac accgccaggt gcacaaccta ccgccccagg
      601 gagcctgaaa gcccctgata cgcgtaacga aaaacttaat tctctggaag acgtacgcaa
      661 aggcagtgaa aattatgcgc tgaccactaa tcagggcgtg cgcatcgccg acgatcaaaa
      721 ctcactgcgt gccggtagcc gtggtccaac gctgctggaa gattttattc tgcgcgagaa
      781 aatcacccac tttgaccatg agcgcattcc ggaacgtatt gttcatgcac gcggatcagc
      841 cgctcacggt tatttccagc catataaaag cttaagcgat attaccaaag cggatttcct
      901 ctcagatccg aacaaaatca ccccagtatt tgtacgtttc tctaccgttc agggtggtgc
      961 tggctctgct gataccgtgc gtgatatccg tggctttgcc accaagttct ataccgaaga
     1021 gggtattttt gacctcgttg gcaataacac gccaatcttc tttatccagg atgcgcataa
     1081 attccccgat tttgttcatg cggtaaaacc agaaccgcac tgggcaattc cacaagggca
     1141 aagtgcccac gatactttct gggattatgt ttctctgcaa cctgaaactc tgcacaacgt
     1201 gatgtgggcg atgtcggatc gcggcatccc ccgcagttac cgcaccatgg aaggcttcgg
     1261 tattcacacc ttccgcctga ttaatgccga agggaaggca acgtttgtac gtttccactg
     1321 gaaaccactg gcaggtaaag cctcactcgt ttgggatgaa gcacaaaaac tcaccggacg
     1381 tgacccggac ttccaccgcc gcgagttgtg ggaagccatt gaagcaggcg attttccgga
     1441 atacgaactg ggcttccagt tgattcctga agaagatgaa ttcaagttcg acttcgatct
     1501 tctcgatcca accaaactta tcccggaaga actggtgccc gttcagcgtg tcggcaaaat
     1561 ggtgctcaat cgcaacccgg ataacttctt tgctgaaaac gaacaggcgg ctttccatcc
     1621 tgggcatatc gtgccgggac tggacttcac caacgatccg ctgttgcagg gacgtttgtt
     1681 ctcctagacc gatacacaaa tcagtcgtct tggtgggccg aatttccatg agattccgat
     1741 taaccgtccg acctgccctt accataattt ccagcgtgac ggcatgcatc gcatggggat
     1801 cgacactaac ccggcgaatt acgaaccgaa ctcgattaac gataactggc cgcgcgaaac
     1861 accgccgggg ccgaaacgcg gcggttttga atcataccag gagcgcgtgg aaggcaataa
     1921 agttcgcgag cgcagcccat cgtttggcga atattattcc catccgcgtc tgttctggct
     1981 aagtcagacg ccatttgagc agcgccatat tgtcgatggt ttcagttttg agttaagcaa
     2041 agtcgttcgt ccgtatattc gtgagcgcgt tgttgaccag ctggcgcata ttgatctcac
     2101 tctggcccag gcggtggcga aaaatctcgg tatcgaactg actgacgacc agctgaatat
     2161 caccccacct ccggacgtca acggtctgaa aaaggatcca tccttaagtt tgtacgccat
     2221 tcctgacggt gatgtgaaag gtcgcgtggt agcgatttta cttaatgatg aagtgagatc
     2281 ggcagacctt ctggccattc tcaaggcgct gaaggccaaa ggcgttcatg ccaaactgct
     2341 ctactcccga atgggtgaag tgactgcgga tgacggtacg gtgttgccta tagccgctac
     2401 ctttgccggt gcaccttcgc tgacggtcga tgcggtcatt gtcccttgcg gcaatatcgc
     2461 ggatatcgct gacaacggcg atgccaacta ctacctgatg gaagcctaca aacaccttaa
     2521 accgattgcg ctggcgggtg acgcgcgcaa gtttaaagca acaatcaaga tcgctgacca
     2581 gggtgaagaa gggattgtgg aagctgacag cgctgacggt agttttatgg atgaactgct
     2641 aacgctgatg gcagcacacc gcgtgtggtc acgcattcct aagattgaca aaattcctgc
     2701 ctgatggggt accatatggg aattcgaagc ttggctgttt tggcggatga gagaagattt
     2761 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     2821 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     2881 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     2941 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     3001 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     3061 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     3121 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     3181 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3241 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3301 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3361 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3421 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3481 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3541 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3601 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3661 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3721 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3781 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3841 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3901 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3961 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4021 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4081 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4141 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4201 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4261 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4321 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4381 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4441 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4501 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4561 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4621 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4681 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4741 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4801 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4861 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4921 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4981 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5041 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5101 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     5161 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     5221 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5281 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag
     5341 atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca
     5401 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat
     5461 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     5521 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     5581 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     5641 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     5701 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     5761 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     5821 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     5881 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     5941 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     6001 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     6061 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     6121 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     6181 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     6241 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     6301 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccg
//
