LOCUS       sequence-337296-        5717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express His6-ExbB S43A, S45A, S67A, S80A, S82A, S84A, S98A, S101A, 
            T111A, S112A, S221A, S229A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  TITLE     Postle Lab Plasmids
REFERENCE   2  (bases 1 to 5717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pKP2032
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1253..1270)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1423..1509
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1601..1628
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1647..1738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1739..1807,1808..2599)
                     /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(1957..1976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2641..3096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2728..2747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2938..2959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3207..3795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3696..3715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3949..3966
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4132..4154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4292..4311
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4491..5369)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5396..5680
                     /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     5603..5622
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 catcaccatc accatcacgg taataattta atgcagacgg acctttccgt ctggggtatg
       61 tatcagcacg ccgatattgt cgttaagtgc gtgatgattg ggcttatttt ggcctccgta
      121 gtcacctggg caatcttctt cgcgaaggca gtagagttct tcaatcagaa gcgtcgcctt
      181 aagcgcgagc agcaactgct ggctgaagcg cgtgctttaa accaggccaa cgatatcgcc
      241 gctgattttg gtgcaaaagc attagcactg catttgctca atgaagcgca gaatgagctg
      301 gaactggcgg aaggcgcgga cgataacgaa ggtattaaag aacgtgcggc attccgtctg
      361 gagcgtcggg tcgccgcagt gggtcgtcaa atgggccgcg gtaacggcta tctggcaacc
      421 atcggcgcga tttctccgtt cgttggtctg tttggtacgg tctggggcat catgaacagc
      481 tttattggta tcgcgcaaac gcagaccact aacctggcag tcgttgcgcc gggtatcgca
      541 gaagctctgt tagcaacggc aatcggcctc gtggcagcga ttcctgcggt cgttatctat
      601 aacgtatttg cacgccagat tggcggcttt aaagcgatgc tgggtgatgt tgcagcgcag
      661 gtattgttgc tgcaagcacg tgacctggat ctggaagccg cagccgctgc gcatccggtt
      721 cgtgtcgcac aaaaattacg cgcaggataa tatccgatgg caatgcatct taacgaaaac
      781 ctcgacgata acggcgaaat gcatgatatc aacgtgacgc cgtttatcga cgtgatgttg
      841 gttctgctga ttatctttat ggtggcggca ccgttagcga cggtagatgt gaaggtgaac
      901 ttgcctgctt ctaccagcac gccgcagccg cgcccggaaa aaccggttta tctgtcggtg
      961 aaggcagaca actcgatgtt tatcggtaac gatccggtca ccgatgaaac aatgattacg
     1021 gcgttgaatg cgttaaccga aggcaagaaa gacaccacca tcttcttccg agcggacaaa
     1081 accgtcgatt acgagacgtt gatgaaggta atggatacgc tgcatcaggc gggttacctg
     1141 aagataggtc tggtcggcga agaaaccgcc aaagcgaagt aaggggatcc tctagagtcg
     1201 acctgcaggc atgcaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     1261 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     1321 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1381 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1441 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1501 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     1561 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     1621 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta
     1681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1981 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2641 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2701 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2761 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2821 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2881 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2941 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     3001 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3061 acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt
     3121 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3181 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3241 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3301 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3361 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3421 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3481 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3541 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3601 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3661 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3721 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3781 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3841 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3901 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3961 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4021 tcacaccgca tagggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc
     4201 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     4261 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     4321 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
     4381 gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg
     4441 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac
     4501 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
     4561 ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga
     4621 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
     4681 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
     4741 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
     4801 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
     4861 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc
     4921 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
     4981 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
     5041 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
     5101 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
     5161 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     5221 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     5281 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     5341 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa
     5401 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
     5461 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
     5521 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
     5581 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct
     5641 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttgggctagc
     5701 aggaggaatt caccatg
//
