LOCUS       sequence-372317-        4610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of heat-resistant obscure (Hero) protein Hero7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4610)
  AUTHORS   Tsuboyama K, Osaki T, Matsuura-Suzuki E, Kozuka-Hata H, Okada Y, 
            Oyama M, Ikeuchi Y, Iwasaki S, Tomari Y
  TITLE     A widespread family of heat-resistant obscure (Hero) proteins 
            protect against protein instability and aggregation.
  JOURNAL   PLoS Biol. 2020 Mar 12;18(3):e3000632. doi: 
            10.1371/journal.pbio.3000632. eCollection 2020 Mar.
  PUBMED    32163402
REFERENCE   2  (bases 1 to 4610)
  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: "PLoS 
            Biol."; date: "2020-03-12"; volume: "18(3):e3000632. doi"; pages: " 
            10.1371/journal.pbio.3000632. eCollection 2020 Mar"
COMMENT     Sequence Label: pCold I-Hero7
FEATURES             Location/Qualifiers
     source          1..4610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..83
                     /gene="cspA"
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    86..102
                     /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)."
     5'UTR           124..257
                     /gene="cspA"
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     CDS             258..272
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="MNHKV"
     CDS             450..473
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             474..491
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             494..505
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     3'UTR           573..717
                     /gene="cspA"
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     primer_bind     complement(758..780)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     880..899
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(916..1371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1053..1074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1265..1284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1471..1575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1576..1644,1645..2436)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1794..1813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2607..3195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3096..3115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3367..4449)
                     /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     4411..4430
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4450..4527)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gcaaggaatg gtgtggccga ttaatcataa atatgaaaaa taattgttgc atcacccgcc
       61 aatgcgtggc ttaatgcaca tcaaattgtg agcggataac aatttgatgt gctagcgcat
      121 atccagtgta gtaaggcaag tcccttcaag agttatcgtt gatacccctc gtagtgcaca
      181 ttcctttaac gcttcaaaat ctgtaaagca cgccatatcg ccgaaaggca cacttaatta
      241 ttaagaggta atacaccatg aatcacaaag tgatgacccg cggtaaccag cgtgagctcg
      301 cccgccagaa gaatatgaaa aagcagagcg actcggttaa gggaaagcgc cgagatgacg
      361 ggctttctgc tgccgcccgc aagcagaggg actcggagat catgcagcag aagcagaaaa
      421 aggcaaacga gaagaaggag gaacccaagg actacaaaga cgatgacgac aagcatcatc
      481 atcatcatca ttaatcgaag gtaggcatat ggagctcggt accctcgagg gatccgaatt
      541 caagcttgtc gacctgcagt ctagataggt aatctctgct taaaagcaca gaatctaaga
      601 tccctgccat ttggcgggga tttttttatt tgttttcagg aaataaataa tcgatcgcgt
      661 aataaaatct attattattt ttgtgaagaa taaatttggg tgcaatgaga atgcgcaggc
      721 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      781 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      841 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      901 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
      961 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1021 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1081 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     1141 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     1201 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1261 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     1321 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
     1381 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1441 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1741 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3241 ccttttgctc acatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     3301 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     3361 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3421 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     3481 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     3541 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     3601 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     3661 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     3721 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     3781 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     3841 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     3901 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     3961 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     4021 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     4081 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     4141 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     4201 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     4261 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     4321 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     4381 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     4441 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     4501 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     4561 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
//
