LOCUS       sequence-298511-        4470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of P. aeruginosa PA14, 6xHis-HRV3C-IHFalpha and IHFbeta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4470)
  AUTHORS   Santiago-Frangos A, Buyukyoruk M, Wiegand T, Krishna P, Wiedenheft B
  TITLE     Distribution and phasing of sequence motifs that facilitate CRISPR 
            adaptation.
  JOURNAL   Curr Biol. 2021 Aug 23;31(16):3515-3524.e6. doi: 
            10.1016/j.cub.2021.05.068. Epub 2021 Jun 25.
  PUBMED    34174210
REFERENCE   2  (bases 1 to 4470)
  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"; doi: 
            "10.1016/j.cub.2021.05.068"; journalName: "Curr Biol"; date: 
            "2021-08-23- 23"; volume: "31"; issue: "16"; pages: "3515-3524.e6"
COMMENT     Sequence Label: pHisIHF-alpha-IHF-beta
FEATURES             Location/Qualifiers
     source          1..4470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..34
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    35..59
                     /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)."
     CDS             115..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             145..168
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        543..561
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    562..586
                     /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)."
     primer_bind     complement(940..958)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      954..1001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1222..1881)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1796..1815
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1882..1984)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2376..2393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2510..3055)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2590..2609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3280..4362)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4324..4343
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4363..4440)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
       61 ctgtagaaat aattttgttt aactttaata aggagatata ccatgggcag cagccatcac
      121 catcatcacc acagccagga tcctctggaa gtgctgtttc agggtccgat gggggctctg
      181 acgaaagctg aaattgcgga acgtctgtac gaagagctgg gcctgaacaa acgggaagcc
      241 aaggagttgg tggagctgtt cttcgaagag atccgccagg cgctggagca caacgagcag
      301 gtcaagctgt ccggtttcgg caacttcgac ctgcgcgaca agcgccagcg ccccggacgc
      361 aacccgaaga ccggtgaaga aatcccgatc acggcccgcc gcgttgtcac ctttcgtcca
      421 gggcagaaac tgaaggccag ggttgaggca tatgctggaa ccaagtcata aaagcttgcg
      481 gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa
      541 attaatacga ctcactatag gggaattgtg agcggataac aattccccat cttagtatat
      601 tagttaagta taagaaggag atatacatat ggcagatcta tgaccaagtc ggagttgatc
      661 gaacggatcg ttacccatca ggggcaactg tccgcgaagg atgtcgagtt ggcaatcaag
      721 accatgctgg agcaaatgtc ccaagccctg gcgactgggg accggatcga gatccgtggc
      781 ttcggcagct tttccttgca ctaccgcgcc ccgcgcgtcg gtcgcaaccc caagaccggg
      841 gagtcggtac gcctcgacgg caagttcgtg ccgcacttca agccgggcaa ggagttgcgg
      901 gatcgggtca acgagccgga gtgacctagg ctgctgccac cgctgagcaa taactagcat
      961 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg
     1021 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
     1081 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
     1141 tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca
     1201 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     1261 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     1321 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatagtga aaacgggggc
     1381 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     1441 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     1501 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     1561 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     1621 actatcccat atcaccagct caccgtcttt cattgccata cggaactccg gatgagcatt
     1681 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     1741 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     1801 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     1861 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     1921 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     1981 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
     2041 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
     2101 gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga ggtgctccag
     2161 tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg gtgcgtaacg
     2221 gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact atgttggcac
     2281 tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt
     2341 cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg
     2401 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     2461 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     2521 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     2581 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     2641 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     2701 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     2761 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     2821 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     2881 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     2941 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     3001 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     3061 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     3121 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     3181 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     3241 gcctaatgag tgagctaact tacattaatt gcattgcgct cactgcccgc tttccagtcg
     3301 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3361 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     3421 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     3481 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     3541 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3601 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3661 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3721 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3781 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     3841 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3901 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3961 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     4021 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     4081 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     4141 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     4201 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     4261 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     4321 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     4381 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     4441 cgggatctcg acgctctccc ttatgcgact
//
