LOCUS       Exported                6354 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses E. coli HPII catalase with TAG stop codon at site 568 and 
            N-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-HPII TAG568
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  JOURNAL   Unpublished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        19..303
                     /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     226..245
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             349..366
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             370..402
                     /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             406..429
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2773..2790)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2943..3029
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3121..3148
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3167..3258
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3259..4119
                     /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(3477..3496)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4290..4878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4779..4798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5032..5049
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5064..5204
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5130..5149)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5290..5312
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5468..6346)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
       61 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
      121 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
      181 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
      241 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
      301 catacccgtt ttttgggcta acaggaggaa ttaaccatgg ggggttctca tcatcatcat
      361 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      421 gacgataagg atcgatgggg atccgagctc gagtcgcaac ataacgaaaa gaacccacat
      481 cagcaccagt caccactaca cgattccagc gaagcgaaac cggggatgga ctcactggca
      541 cctgaggacg gctctcatcg tccagcggct gaaccaacac cgccaggtgc acaacctacc
      601 gccccaggga gcctgaaagc ccctgatacg cgtaacgaaa aacttaattc tctggaagac
      661 gtacgcaaag gcagtgaaaa ttatgcgctg accactaatc agggcgtgcg catcgccgac
      721 gatcaaaact cactgcgtgc cggtagccgt ggtccaacgc tgctggaaga ttttattctg
      781 cgcgagaaaa tcacccactt tgaccatgag cgcattccgg aacgtattgt tcatgcacgc
      841 ggatcagccg ctcacggtta tttccagcca tataaaagct taagcgatat taccaaagcg
      901 gatttcctct cagatccgaa caaaatcacc ccagtatttg tacgtttctc taccgttcag
      961 ggtggtgctg gctctgctga taccgtgcgt gatatccgtg gctttgccac caagttctat
     1021 accgaagagg gtatttttga cctcgttggc aataacacgc caatcttctt tatccaggat
     1081 gcgcataaat tccccgattt tgttcatgcg gtaaaaccag aaccgcactg ggcaattcca
     1141 caagggcaaa gtgcccacga tactttctgg gattatgttt ctctgcaacc tgaaactctg
     1201 cacaacgtga tgtgggcgat gtcggatcgc ggcatccccc gcagttaccg caccatggaa
     1261 ggcttcggta ttcacacctt ccgcctgatt aatgccgaag ggaaggcaac gtttgtacgt
     1321 ttccactgga aaccactggc aggtaaagcc tcactcgttt gggatgaagc acaaaaactc
     1381 accggacgtg acccggactt ccaccgccgc gagttgtggg aagccattga agcaggcgat
     1441 tttccggaat acgaactggg cttccagttg attcctgaag aagatgaatt caagttcgac
     1501 ttcgatcttc tcgatccaac caaacttatc ccggaagaac tggtgcccgt tcagcgtgtc
     1561 ggcaaaatgg tgctcaatcg caacccggat aacttctttg ctgaaaacga acaggcggct
     1621 ttccatcctg ggcatatcgt gccgggactg gacttcacca acgatccgct gttgcaggga
     1681 cgtttgttct cctataccga tacacaaatc agtcgtcttg gtgggccgaa tttccatgag
     1741 attccgatta accgtccgac ctgcccttac cataatttcc agcgtgacgg catgcatcgc
     1801 atggggatcg acactaaccc ggcgaattac gaaccgaact cgattaacga taactggccg
     1861 cgcgaaacac cgccggggcc gaaacgcggc ggttttgaat cataccagga gcgcgtggaa
     1921 ggcaataaag ttcgcgagcg cagcccatcg tttggcgaat attattccca tccgcgtctg
     1981 ttctggctaa gtcagacgcc atttgagcag cgccatattg tcgatggttt cagttttgag
     2041 ttaagcaaag tcgttcgtcc gtatattcgt gagcgcgttg ttgaccagct ggcgcatatt
     2101 gatctcactc tggcccaggc ggtggcgaaa aatctcggta tcgaactgac ttaggaccag
     2161 ctgaatatca ccccacctcc ggacgtcaac ggtctgaaaa aggatccatc cttaagtttg
     2221 tacgccattc ctgacggtga tgtgaaaggt cgcgtggtag cgattttact taatgatgaa
     2281 gtgagatcgg cagaccttct ggccattctc aaggcgctga aggccaaagg cgttcatgcc
     2341 aaactgctct actcccgaat gggtgaagtg actgcggatg acggtacggt gttgcctata
     2401 gccgctacct ttgccggtgc accttcgctg acggtcgatg cggtcattgt cccttgcggc
     2461 aatatcgcgg atatcgctga caacggcgat gccaactact acctgatgga agcctacaaa
     2521 caccttaaac cgattgcgct ggcgggtgac gcgcgcaagt ttaaagcaac aatcaagatc
     2581 gctgaccagg gtgaagaagg gattgtggaa gctgacagcg ctgacggtag ttttatggat
     2641 gaactgctaa cgctgatggc agcacaccgc gtgtggtcac gcattcctaa gattgacaaa
     2701 attcctgcct gatggggtac catatgggaa ttcgaagctt ggctgttttg gcggatgaga
     2761 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     2821 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     2881 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     2941 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3001 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     3061 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     3121 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     3181 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3241 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3301 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3361 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3421 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3481 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     3541 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3601 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3661 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3721 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3781 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3841 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3901 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3961 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4021 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4081 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4141 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4201 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4261 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4321 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4381 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4441 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4501 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4561 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4621 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4681 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4741 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4801 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4861 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4921 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4981 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5041 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5101 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5161 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     5221 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5281 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5341 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg
     5401 acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg
     5461 ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     5521 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     5581 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     5641 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     5701 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     5761 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     5821 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     5881 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     5941 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     6001 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     6061 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     6121 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     6181 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     6241 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     6301 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttca
//
