LOCUS       sequence-360535-        4184 bp    DNA     circular SYN 15-APR-2025
DEFINITION  E. coli plasmid encoding pH-Countdown with a hexahistidine tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4184)
  AUTHORS   Seidel ZP, Wang JCK, Riegler J, York AG, Ingaramo M
  TITLE     Relaxation Sensors
  JOURNAL   (2021), doi: 10.5281/zenodo.5810930
REFERENCE   2  (bases 1 to 4184)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "(2021), 
            doi: 10.5281/zenodo.5810930"
COMMENT     Sequence Label: pQE31-6xHis-pHCountdown
FEATURES             Location/Qualifiers
     source          1..4184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      932..1026
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1136..1155)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1793..1879
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1926..1948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2034..2174
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2089..2108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2189..2206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2360..2948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2440..2459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3119..3910,3911..3979))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNNGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASWQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3742..3761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3980..4084)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4152..4170
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatacggat ccaatggtga gcaagggcga ggagaacaac
      181 atggccgtga ttaaaccaga catgaagatc aagctgcgta tggaaggctc tgtaaatgga
      241 caccgtttca gaattgaagg agttggcctt gggaagcctc ttgagggaaa acagagtatg
      301 gaccttaaag tcaaagaagg cggacctctg cctttcgcct atgacatctt gacaatggcg
      361 ttctgttacg gtaacagggt attcgccaaa tacccagaaa atatagtaga ctatttcaag
      421 cagtcgtttc ctgagggcta ctcttgggaa cgacaaatga tttacgaaga cgggggcatt
      481 tgtgtcgcga caaacgacat aaccctggat ggtgactgta tgatctccga aattcgattt
      541 aagggtgtga actttcctgc caatggtcca gttttccaga agaggactgt gaaatgggag
      601 ctatcccacg agaaattgta tgctcgtgat ggactgctgt actctgatgg taactacgct
      661 ctgtcgcttg aaggaggtgg ccattaccga tgtgacaata aaactactta taaagctaag
      721 aaggttgtcc agttgccaga ctatcactgg gtgacccaca gtattgtgat taaaagccac
      781 gacaaagatt acagtaatgt taatctgcat gagcatgccg aagcacattc tgagctgccg
      841 aggcaggcca tggacgagct gtacaagtaa gaattcgaat ctatcaacag gagtccaagc
      901 tcagctaatt aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
      961 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     1021 gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1081 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1141 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1201 tttaaagacc gtaaagaaaa ataagcacaa gttttattcg gcctttattc acattcttgc
     1261 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     1321 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1381 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcagatgtg
     1441 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1501 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1561 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1621 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1681 cttaatgaat tacaacagta ctgcgatgag tggcagggca gggcgtaatt tttttaaggc
     1741 agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa
     1801 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1861 tctcctgagt aggacaaatc cgccctctag agctgcctcg cgcgtttcgg tgatgacggt
     1921 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1981 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     2041 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     2101 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2161 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2221 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2281 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2521 cctttctccc ttcgggaagc atggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccaacc
     2641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctatag
     2761 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3361 ctattaattg ttgccaggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgttg ttgagatcca
     3841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4021 attgtcgcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4081 cgcgcacatt tcctcgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4141 taacctataa aaataggcgt atcacgaggc cctttcgtct tcac
//
