LOCUS       sequence-420518-        5752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human C17orf53 N-terminally fused to a 
            6xHis-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5752)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5752)
  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"
COMMENT     Sequence Label: pRSF-Duet-6His-C17orf53
FEATURES             Location/Qualifiers
     source          1..5752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    19..43
                     /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             99..116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2153..2171
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2172..2196
                     /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             2305..2349
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2387..2405)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2401..2448
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2678..3493)
                     /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(2678..3493)
                     /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     3401..3420
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3494..3585)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3601..4350)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3611..3628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3851..3870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4546..5628)
                     /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     5590..5609
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5629..5706)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5752,1..18)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aatacgactc actatagggg aattgtgagc ggataacaat tcccctgtag aaataatttt
       61 gtttaacttt aataaggaga tataccatgg gcagcagcca tcaccatcat caccacagcc
      121 aggatccgaa ttctatggcg tgcagtttgc agaagctgtt tgctgtggaa gaggagtttg
      181 aagatgagga tttcttgtct gctgtggagg atgcagagaa ccggtttact ggctcactgc
      241 ctgtgaatgc tgggcgcctg agacctgtct cttctaggcc acaggagact gtgcaggcac
      301 agtcctccag gctgctgctg ttacacccca ctgctccctc agaggctttg ggcctgccag
      361 acttggacct ctgcctccct gcctccagca cgcccagtgc tgacagccgt ccatcatgca
      421 taggagcagc tcccctaagg cctgtctcta cttccagcag ctggattggc aatcagagaa
      481 gagtgacagt gacagaagtg ctcagagaga cagcaagacc tcagtcctca gccttacacc
      541 ccctactcac ctttgagagc caacagcagc aagttggtgg ctttgagggg cctgaacaag
      601 acgaatttga taaagtcctg gcaagcatgg agttggagga gcctggcatg gagctggaat
      661 gtggagtcag cagtgaggcc ataccaatcc tgcctgccca gcagcgggag ggttcagtat
      721 tggctaaaaa agcccgggta gttgatctga gtggatcttg ccagaagggg cctgtgcctg
      781 ccatccacaa agcgggtatc atgtccgccc aggatgagtc tctagatcct gtcatccaat
      841 gtaggactcc acgacccccc ttgagacctg gtgctgtggg tcaccttcct gttccaactg
      901 ccttaacagt tcccactcag caactccact gggaagtctg tccgcaacgc tcccctgttc
      961 aagcacttca gcctctccaa gctgctagag ggaccattca gagcagccct caaaatcgtt
     1021 tcccttgtca gccattccag tctccaagtt cctggttaag tggcaaagct catttaccca
     1081 gacctcgaac tcccaactca agctgttcta ctccctcaag gactagctct ggattatttc
     1141 ctcggatacc cttacaaccg caagctccag tgtcttccat tgggtctcct gttggtaccc
     1201 caaaaggtcc ccagggagct ctgcagacac ccatagtcac caaccacctg gtgcagctag
     1261 tcactgctgc cagccggaca ccccagcagc ccacccatcc ctccacccga gccaaaactc
     1321 gccgtttccc tggcccagct gggatcctgc ctcaccagca gagtgggaga agtctggagg
     1381 acatcatggt ttccgcgccc caaactccaa cccatggtgc tctggctaaa ttccagacag
     1441 agattgttgc tagttcccag gcatctgtgg aggaggattt tgggcgaggg ccctggctga
     1501 ccatgaaatc cacgctaggc ctggatgaga gagaccctag ctgcttcctc tgtacctaca
     1561 gcattgtcat ggtgctgcgc aaggcagccc tgaagcagct tcctaggaac aaggtcccca
     1621 acatggcggt gatgatcaag tccctgactc ggagcacaat ggacgccagt gtggttttca
     1681 aggaccccac gggagagatg caggggacgg tgcacaggtt gctgctggag acgtgccaga
     1741 atgagctgaa gcctggctca gtgctgctgc tgaagcagat tggagtgttt tctccttcac
     1801 ttcgaaatca ctacctcaac gtgacaccca acaacctggt ccatatttac agcccggatt
     1861 ctggggatgg gagcttcctc aagccatctc agcccttccc caaggattca gggagcttcc
     1921 agcatgatgt ggctgcaaag cccgagaaag gcttcagaac agcacagaac ctagaggcag
     1981 aggcgtcccc tgaggaagaa ctcccagaag cagatgacct ggatggactc ctgagtgagc
     2041 ttcctgaaga cttcttctgt gggaccagta gttgagtcga caagcttgcg gccgcataat
     2101 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
     2161 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
     2221 taagaaggag atatacatat ggcagatctc aattggatat cggccggcca cgcgatcgct
     2281 gacgtcggta ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
     2341 atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa
     2401 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     2461 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     2521 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt
     2581 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2641 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     2701 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     2761 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     2821 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     2881 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     2941 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3001 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3061 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3121 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3181 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3241 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     3301 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     3361 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3421 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     3481 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgcagcgct
     3601 cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt
     3661 cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga
     3721 acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg
     3781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg
     3841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3961 tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt
     4141 agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg
     4201 tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt
     4261 gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg
     4321 tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc
     4381 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     4441 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4501 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4561 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4621 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4681 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4741 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4801 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     4861 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4921 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4981 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5041 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5101 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5161 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5221 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5281 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5341 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5401 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5461 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5521 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5581 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5641 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5701 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa tt
//
