LOCUS       sequence-169442-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express E. coli ribosomal protein S1 with a TEV-cleavable, 
            N-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Lund PE, Chatterjee S, Daher M, Walter NG
  TITLE     Protein unties the pseudoknot: S1-mediated unfolding of RNA higher 
            order structure.
  JOURNAL   Nucleic Acids Res. 2019 Dec 13. pii: 5674991. doi: 
            10.1093/nar/gkz1166.
  PUBMED    31832686
REFERENCE   2  (bases 1 to 6194)
  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"; journalName: "Nucleic 
            Acids Res. 2019 Dec 13. pii: 5674991. doi: 10.1093/nar/gkz1166."
COMMENT     Sequence Label: pCA24N_6xHis-TEV-rpsA
FEATURES             Location/Qualifiers
     source          1..6194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     819..836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        909..986
                     /gene="lacI"
                     /label=lacI promoter
     CDS             987..2069
                     /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     complement(1006..1025)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2118..2123,2124..2141,2142..2148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2156..2172
                     /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     2180..2196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2180..2196
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2212..2229)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2212..2228)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2221..2243)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2428..2442,2443..2448,2449..2465,2466..2472)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2448..2464
                     /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    2480..2496
                     /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             2515..2526
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2545..2562
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2563..2583
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      4307..4401
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4471..4493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4958..5060
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5061..5720
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5127..5146)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      901 taaattccga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
      961 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     1021 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     1081 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     1141 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     1201 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     1261 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     1321 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     1381 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     1441 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     1501 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     1561 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     1621 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     1681 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     1741 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     1801 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     1861 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     1921 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     1981 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2041 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact
     2101 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2161 agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt
     2221 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2281 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2341 acagttgcct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2401 aggccctttc gtcttcacct cgagaaatca taaaaaattt atttgctttg tgagcggata
     2461 acaattataa tagattcaat tgtgagcgga taacaatttc acacagaatt cattaaagag
     2521 gagaaattaa ctatgagagg atctcaccat caccatcacc atgagaatct gtattttcag
     2581 ggcactgaat cttttgctca actctttgaa gagtccttaa aagaaatcga aacccgcccg
     2641 ggttctatcg ttcgtggcgt tgttgttgct atcgacaaag acgtagtact ggttgacgct
     2701 ggtctgaaat ctgagtccgc catcccggct gagcagttca aaaacgccca gggcgagctg
     2761 gaaatccagg taggtgacga agttgacgtt gctctggacg cagtagaaga cggcttcggt
     2821 gaaactctgc tgtcccgtga gaaagctaaa cgtcacgaag cctggatcac gctggaaaaa
     2881 gcttacgaag atgctgaaac tgttaccggt gttatcaacg gcaaagttaa gggcggcttc
     2941 actgttgagc tgaacggtat tcgtgcgttc ctgccaggtt ctctggtaga cgttcgtccg
     3001 gtgcgtgaca ctctgcacct ggaaggcaaa gagcttgaat ttaaagtaat caagctggat
     3061 cagaagcgca acaacgttgt tgtttctcgt cgtgccgtta tcgaatccga aaacagcgca
     3121 gagcgcgatc agctgctgga aaacctgcag gaaggcatgg aagttaaagg tatcgttaag
     3181 aacctcactg actacggtgc attcgttgat ctgggcggcg ttgacggcct gctgcacatc
     3241 actgacatgg cctggaaacg cgttaagcat ccgagcgaaa tcgtcaacgt gggcgacgaa
     3301 atcactgtta aagtgctgaa gttcgaccgc gaacgtaccc gtgtatccct gggcctgaaa
     3361 cagctgggcg aagatccgtg ggtagctatc gctaaacgtt atccggaagg taccaaactg
     3421 actggtcgcg tgaccaacct gaccgactac ggctgcttcg ttgaaatcga agaaggcgtt
     3481 gaaggcctgg tacacgtttc cgaaatggac tggaccaaca aaaacatcca cccgtccaaa
     3541 gttgttaacg ttggcgatgt agtggaagtt atggttctgg atatcgacga agaacgtcgt
     3601 cgtatctccc tgggtctgaa acagtgcaaa gctaacccgt ggcagcagtt cgcggaaacc
     3661 cacaacaagg gcgaccgtgt tgaaggtaaa atcaagtcta tcactgactt cggtatcttc
     3721 atcggcttgg acggcggcat cgacggcctg gttcacctgt ctgacatctc ctggaacgtt
     3781 gcaggcgaag aagcagttcg tgaatacaaa aaaggcgacg aaatcgctgc agttgttctg
     3841 caggttgacg cagaacgtga acgtatctcc ctgggcgtta aacagctcgc agaagatccg
     3901 ttcaacaact gggttgctct gaacaagaaa ggcgctatcg taaccggtaa agtaactgca
     3961 gttgacgcta aaggcgcaac cgtagaactg gctgacggcg ttgaaggtta cctgcgtgct
     4021 tctgaagcat cccgtgaccg cgttgaagac gctaccctgg ttctgagcgt tggcgacgaa
     4081 gttgaagcta aattcaccgg cgttgatcgt aaaaaccgcg caatcagcct gtctgttcgt
     4141 gcgaaagacg aagctgacga gaaagatgca atcgcaactg ttaacaaaca ggaagatgca
     4201 aacttctcca acaacgcaat ggctgaagct ttcaaagcag ctaaaggcga gtaactatgc
     4261 ggccgctaag ggtcgacctg cagccaagct taattagctg agcttggact cctgttgata
     4321 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     4381 gcgtttttta ttggtgagaa tccaagctag gggaattcat cgtgactgac tgacgatctg
     4441 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4501 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4561 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa
     4621 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4681 aatggtttct taggatccgg gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc
     4741 agtagctgaa caggagggac agctgataga aacagaagcc actggagcac ctcaaaaaca
     4801 ccatcataca ctaaatcagt aagttggcag catcacccga cgcactttgc gccgaataaa
     4861 tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg
     4921 ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca
     4981 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     5041 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     5101 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     5161 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     5221 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     5281 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     5341 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     5401 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     5461 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     5521 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     5581 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     5641 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     5701 agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt
     5761 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
     5821 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
     5881 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt
     5941 tgctcaggct ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca
     6001 gagcctgata aaaacggtta ccggatcgag tcaggcaact atggatgaac gaaatagaca
     6061 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6121 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6181 cctttttgat aatc
//
