LOCUS       sequence-420530-        3895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of the N-terminal amino acids 1-87 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 3895)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 3895)
  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_1-87-Strep
FEATURES             Location/Qualifiers
     source          1..3895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5..23
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    24..48
                     /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             353..376
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             453..497
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(535..553)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      549..596
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(826..1641)
                     /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(826..1641)
                     /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     1549..1568
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1642..1733)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1749..2498)
                     /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(1759..1776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1999..2018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2694..3776)
                     /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     3738..3757
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3777..3854)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata
       61 attttgttta actttaataa ggagatatac catggcgtgc agtttgcaga agctgtttgc
      121 tgtggaagag gagtttgaag atgaggattt cttgtctgct gtggaggatg cagagaaccg
      181 gtttactggc tcactgcctg tgaatgctgg gcgcctgaga cctgtctctt ctaggccaca
      241 ggagactgtg caggcacagt cctccaggct gctgctgtta caccccactg ctccctcaga
      301 ggctttgggc ctgccagact tggacctctg cctccctgcc tccagcacgc cctggagcca
      361 cccgcagttc gaaaaatgac atatgtatgg cagatctcaa ttggatatcg gccggccacg
      421 cgatcgctga cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac
      481 gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg
      541 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      601 aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
      661 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac
      721 gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      781 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
      841 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
      901 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
      961 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     1021 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     1081 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     1141 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     1201 aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     1261 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     1321 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     1381 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     1441 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     1501 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     1561 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     1621 ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat
     1741 gcagcgctct tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     1801 agcggtgtca gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc
     1861 cggaaagaac atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt
     1921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc
     1981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2101 tggcgctttc tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca
     2161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta
     2281 actgatttag aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca
     2341 gattttggtg agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag
     2401 cgaaccttga gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa
     2461 tcaatcggtc tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt
     2521 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
     2581 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     2641 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2761 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2821 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2881 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     2941 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     3001 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3061 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3121 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3181 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3241 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     3301 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3361 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3421 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3481 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3541 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     3601 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3661 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3721 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3781 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3841 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttagg
//
