LOCUS       sequence-352517-        5500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a peptide (reside from 550 through 600) of human ELMSAN1 
            (Q6PJG2) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5500)
  AUTHORS   Iuchi S, Paulo JA
  TITLE     RNAmetasome network for macromolecule biogenesis in human cells.
  JOURNAL   Commun Biol. 2021 Dec 15;4(1):1399. doi: 10.1038/s42003-021-02928-y.
  PUBMED    34912035
REFERENCE   2  (bases 1 to 5500)
  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: "Commun 
            Biol."; date: "2021-12-15"; volume: "4(1):1399. doi"; pages: " 
            10.1038/s42003-021-02928-y"
COMMENT     Sequence Label: pET-epELM-5
FEATURES             Location/Qualifiers
     source          1..5500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     925..944
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(964..1041)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1247..1266
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1350..1369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1350..1368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1369..1393
                     /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             1408..1430
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1449..1466
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1476..1493
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1497..1529
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1714..1731
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1784..1802)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1798..1845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1882..2337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1969..1988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2179..2200
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2429..3244)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2429..3244)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3152..3171
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3236..3255
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3366..3954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3855..3874
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4108..4125
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4140..4279
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4205..4224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4365..4387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4381..4572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5381..5500,1..963))
                     /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"
ORIGIN
        1 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
       61 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
      121 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
      181 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
      241 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
      301 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
      361 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
      421 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
      481 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
      541 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
      601 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
      661 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
      721 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
      781 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
      841 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
      901 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
      961 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     1021 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     1081 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1141 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1201 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1261 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1321 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     1381 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
     1441 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgg
     1501 ctagcatgac tggtggacag caaatgggtc ggatccttga cgggaagggt cctgaacaga
     1561 accctgctga gcacaagcca tcagtcatcg tcacccgcag gcggtccacc cgaatccccg
     1621 ggacagatgc tcaagctcag gcagaggaca tgaatgtcaa gttggagggg gagccttccg
     1681 tgcggaaacc ataagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg
     1741 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1801 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1861 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1981 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2041 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2101 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2161 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2221 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2281 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     2341 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2401 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
     2461 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2521 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     2581 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2641 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     2701 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2761 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     2821 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     2881 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     2941 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3001 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3061 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     3121 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3181 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     3241 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa
     3301 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3361 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3421 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3481 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3541 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3601 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3661 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3721 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3781 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3841 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3901 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3961 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4021 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4081 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4141 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
     4201 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     4261 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4321 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4381 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     4441 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4501 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4561 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     4621 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     4681 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     4741 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     4801 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     4861 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     4921 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     4981 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     5041 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     5101 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     5161 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     5221 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     5281 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     5341 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5401 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5461 gcgtattggg cgccagggtg gtttttcttt tcaccagtga
//
