LOCUS       sequence-420531-        4351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of the N-terminal amino acids 1-241 of human 
            C17orf53 C-terminally fused to a Strep-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4351)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 4351)
  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-C17orf53_1-241-Strep
FEATURES             Location/Qualifiers
     source          1..4351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..26
                     /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             793..816
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             887..931
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(969..987)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      983..1030
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1260..2075)
                     /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(1260..2075)
                     /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     1983..2002
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2076..2167)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2183..2932)
                     /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(2193..2210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2433..2452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3128..4210)
                     /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     4172..4191
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4211..4288)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4334..4351,1..2)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4334..4351,1)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggaattgtg agcggataac aattcccctg tagaaataat tttgtttaac tttaataagg
       61 agatatacca tggcgtgcag tttgcagaag ctgtttgctg tggaagagga gtttgaagat
      121 gaggatttct tgtctgctgt ggaggatgca gagaaccggt ttactggctc actgcctgtg
      181 aatgctgggc gcctgagacc tgtctcttct aggccacagg agactgtgca ggcacagtcc
      241 tccaggctgc tgctgttaca ccccactgct ccctcagagg ctttgggcct gccagacttg
      301 gacctctgcc tccctgcctc cagcacgccc agtgctgaca gccgtccatc atgcatagga
      361 gcagctcccc taaggcctgt ctctacttcc agcagctgga ttggcaatca gagaagagtg
      421 acagtgacag aagtgctcag agagacagca agacctcagt cctcagcctt acacccccta
      481 ctcacctttg agagccaaca gcagcaagtt ggtggctttg aggggcctga acaagacgaa
      541 tttgataaag tcctggcaag catggagttg gaggagcctg gcatggagct ggaatgtgga
      601 gtcagcagtg aggccatacc aatcctgcct gcccagcagc gggagggttc agtattggct
      661 aaaaaagccc gggtagttga tctgagtgga tcttgccaga aggggcctgt gcctgccatc
      721 cacaaagcgg gtatcatgtc cgcccaggat gagtctctag atcctgtcat ccaatgtagg
      781 actccacgac cctggagcca cccgcagttc gaaaaatgac atggcagatc tcaattggat
      841 atcggccggc cacgcgatcg ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc
      901 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc
      961 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1021 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     1081 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     1141 gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat gtgcgcggaa
     1201 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     1261 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1321 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1381 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1441 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1501 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1561 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1621 gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac aattacaaac aggaatcgaa
     1681 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     1741 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     1801 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     1861 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     1921 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     1981 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     2041 ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga
     2101 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2161 aaacaaatag gcatgcagcg ctcttccgct tcctcgctca ctgactcgct acgctcggtc
     2221 gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg taatacggtt atccacagaa
     2281 tcaggggata aagccggaaa gaacatgtga gcaaaaagca aagcaccgga agaagccaac
     2341 gccgcaggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2401 aagccagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt tggtatctca gttcggtgta
     2581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2701 agcagccatt ggtaactgat ttagaggact ttgtcttgaa gttatgcacc tgttaaggct
     2761 aaactgaaag aacagatttt ggtgagtgcg gtcctccaac ccacttacct tggttcaaag
     2821 agttggtagc tcagcgaacc ttgagaaaac caccgttggt agcggtggtt tttctttatt
     2881 tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac agctattccg ttactctaga
     2941 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     3001 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     3061 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     3121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     3241 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     3301 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     3361 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     3421 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     3481 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     3541 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     3601 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     3661 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     3721 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     3781 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     3841 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     3901 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     3961 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     4021 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     4081 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     4141 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     4201 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     4261 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     4321 tgcattagga aattaatacg actcactata g
//
