LOCUS       Exported                6354 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses E. coli HPII catalase with TAG stop codon at site 206 and 
            N-terminal 6x his tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBad-HPII TAG206
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  JOURNAL   Unpublished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        12..296
                     /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     219..238
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             342..359
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             363..395
                     /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             399..422
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2766..2783)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2936..3022
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3114..3141
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3160..3251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3252..4112
                     /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(3470..3489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4283..4871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4772..4791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5025..5042
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5057..5197
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5123..5142)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5283..5305
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5461..6339)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
       61 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
      121 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
      181 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
      241 aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
      301 gttttttggg ctaacaggag gaattaacca tggggggttc tcatcatcat catcatcatg
      361 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      421 aggatcgatg gggatccgag ctcgagtcgc aacataacga aaagaaccca catcagcacc
      481 agtcaccact acacgattcc agcgaagcga aaccggggat ggactcactg gcacctgagg
      541 acggctctca tcgtccagcg gctgaaccaa caccgccagg tgcacaacct accgccccag
      601 ggagcctgaa agcccctgat acgcgtaacg aaaaacttaa ttctctggaa gacgtacgca
      661 aaggcagtga aaattatgcg ctgaccacta atcagggcgt gcgcatcgcc gacgatcaaa
      721 actcactgcg tgccggtagc cgtggtccaa cgctgctgga agattttatt ctgcgcgaga
      781 aaatcaccca ctttgaccat gagcgcattc cggaacgtat tgttcatgca cgcggatcag
      841 ccgctcacgg ttatttccag ccatataaaa gcttaagcga tattaccaaa gcggatttcc
      901 tctcagatcc gaacaaaatc accccagtat ttgtacgttt ctctaccgtt cagggtggtg
      961 ctggctctgc tgataccgtg cgtgatatcc gtggctttgc caccaagttc tataccgaag
     1021 agggtatttt tgacctcgtt ggcaataaca cgccaatcta gtttatccag gatgcgcata
     1081 aattccccga ttttgttcat gcggtaaaac cagaaccgca ctgggcaatt ccacaagggc
     1141 aaagtgccca cgatactttc tgggattatg tttctctgca acctgaaact ctgcacaacg
     1201 tgatgtgggc gatgtcggat cgcggcatcc cccgcagtta ccgcaccatg gaaggcttcg
     1261 gtattcacac cttccgcctg attaatgccg aagggaaggc aacgtttgta cgtttccact
     1321 ggaaaccact ggcaggtaaa gcctcactcg tttgggatga agcacaaaaa ctcaccggac
     1381 gtgacccgga cttccaccgc cgcgagttgt gggaagccat tgaagcaggc gattttccgg
     1441 aatacgaact gggcttccag ttgattcctg aagaagatga attcaagttc gacttcgatc
     1501 ttctcgatcc aaccaaactt atcccggaag aactggtgcc cgttcagcgt gtcggcaaaa
     1561 tggtgctcaa tcgcaacccg gataacttct ttgctgaaaa cgaacaggcg gctttccatc
     1621 ctgggcatat cgtgccggga ctggacttca ccaacgatcc gctgttgcag ggacgtttgt
     1681 tctcctatac cgatacacaa atcagtcgtc ttggtgggcc gaatttccat gagattccga
     1741 ttaaccgtcc gacctgccct taccataatt tccagcgtga cggcatgcat cgcatgggga
     1801 tcgacactaa cccggcgaat tacgaaccga actcgattaa cgataactgg ccgcgcgaaa
     1861 caccgccggg gccgaaacgc ggcggttttg aatcatacca ggagcgcgtg gaaggcaata
     1921 aagttcgcga gcgcagccca tcgtttggcg aatattattc ccatccgcgt ctgttctggc
     1981 taagtcagac gccatttgag cagcgccata ttgtcgatgg tttcagtttt gagttaagca
     2041 aagtcgttcg tccgtatatt cgtgagcgcg ttgttgacca gctggcgcat attgatctca
     2101 ctctggccca ggcggtggcg aaaaatctcg gtatcgaact gactgacgac cagctgaata
     2161 tcaccccacc tccggacgtc aacggtctga aaaaggatcc atccttaagt ttgtacgcca
     2221 ttcctgacgg tgatgtgaaa ggtcgcgtgg tagcgatttt acttaatgat gaagtgagat
     2281 cggcagacct tctggccatt ctcaaggcgc tgaaggccaa aggcgttcat gccaaactgc
     2341 tctactcccg aatgggtgaa gtgactgcgg atgacggtac ggtgttgcct atagccgcta
     2401 cctttgccgg tgcaccttcg ctgacggtcg atgcggtcat tgtcccttgc ggcaatatcg
     2461 cggatatcgc tgacaacggc gatgccaact actacctgat ggaagcctac aaacacctta
     2521 aaccgattgc gctggcgggt gacgcgcgca agtttaaagc aacaatcaag atcgctgacc
     2581 agggtgaaga agggattgtg gaagctgaca gcgctgacgg tagttttatg gatgaactgc
     2641 taacgctgat ggcagcacac cgcgtgtggt cacgcattcc taagattgac aaaattcctg
     2701 cctgatgggg taccatatgg gaattcgaag cttggctgtt ttggcggatg agagaagatt
     2761 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     2821 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     2881 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     2941 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     3001 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     3061 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     3121 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     3181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3481 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4921 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4981 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5041 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5101 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     5161 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     5221 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5281 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
     5341 gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc
     5401 agggattctg caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa
     5461 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
     5521 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
     5581 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
     5641 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
     5701 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
     5761 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
     5821 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
     5881 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
     5941 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
     6001 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
     6061 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
     6121 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
     6181 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
     6241 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
     6301 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tccc
//
