LOCUS       sequence-420520-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human C17orf53 C-terminally fused to 
            6xHis-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5513)
  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-C17orf53-6His
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    22..46
                     /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             2036..2053
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2069..2113
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2151..2169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2165..2212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2442..3257)
                     /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(2442..3257)
                     /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     3165..3184
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3258..3349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3365..4114)
                     /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(3375..3392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4310..5392)
                     /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     5354..5373
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5393..5470)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 attaatacga ctcactatag gggaattgtg agcggataac aattcccctg tagaaataat
       61 tttgtttaac tttaataagg agatatacca tgttatggcg tgcagtttgc agaagctgtt
      121 tgctgtggaa gaggagtttg aagatgagga tttcttgtct gctgtggagg atgcagagaa
      181 ccggtttact ggctcactgc ctgtgaatgc tgggcgcctg agacctgtct cttctaggcc
      241 acaggagact gtgcaggcac agtcctccag gctgctgctg ttacacccca ctgctccctc
      301 agaggctttg ggcctgccag acttggacct ctgcctccct gcctccagca cgcccagtgc
      361 tgacagccgt ccatcatgca taggagcagc tcccctaagg cctgtctcta cttccagcag
      421 ctggattggc aatcagagaa gagtgacagt gacagaagtg ctcagagaga cagcaagacc
      481 tcagtcctca gccttacacc ccctactcac ctttgagagc caacagcagc aagttggtgg
      541 ctttgagggg cctgaacaag acgaatttga taaagtcctg gcaagcatgg agttggagga
      601 gcctggcatg gagctggaat gtggagtcag cagtgaggcc ataccaatcc tgcctgccca
      661 gcagcgggag ggttcagtat tggctaaaaa agcccgggta gttgatctga gtggatcttg
      721 ccagaagggg cctgtgcctg ccatccacaa agcgggtatc atgtccgccc aggatgagtc
      781 tctagatcct gtcatccaat gtaggactcc acgacccccc ttgagacctg gtgctgtggg
      841 tcaccttcct gttccaactg ccttaacagt tcccactcag caactccact gggaagtctg
      901 tccgcaacgc tcccctgttc aagcacttca gcctctccaa gctgctagag ggaccattca
      961 gagcagccct caaaatcgtt tcccttgtca gccattccag tctccaagtt cctggttaag
     1021 tggcaaagct catttaccca gacctcgaac tcccaactca agctgttcta ctccctcaag
     1081 gactagctct ggattatttc ctcggatacc cttacaaccg caagctccag tgtcttccat
     1141 tgggtctcct gttggtaccc caaaaggtcc ccagggagct ctgcagacac ccatagtcac
     1201 caaccacctg gtgcagctag tcactgctgc cagccggaca ccccagcagc ccacccatcc
     1261 ctccacccga gccaaaactc gccgtttccc tggcccagct gggatcctgc ctcaccagca
     1321 gagtgggaga agtctggagg acatcatggt ttccgcgccc caaactccaa cccatggtgc
     1381 tctggctaaa ttccagacag agattgttgc tagttcccag gcatctgtgg aggaggattt
     1441 tgggcgaggg ccctggctga ccatgaaatc cacgctaggc ctggatgaga gagaccctag
     1501 ctgcttcctc tgtacctaca gcattgtcat ggtgctgcgc aaggcagccc tgaagcagct
     1561 tcctaggaac aaggtcccca acatggcggt gatgatcaag tccctgactc ggagcacaat
     1621 ggacgccagt gtggttttca aggaccccac gggagagatg caggggacgg tgcacaggtt
     1681 gctgctggag acgtgccaga atgagctgaa gcctggctca gtgctgctgc tgaagcagat
     1741 tggagtgttt tctccttcac ttcgaaatca ctacctcaac gtgacaccca acaacctggt
     1801 ccatatttac agcccggatt ctggggatgg gagcttcctc aagccatctc agcccttccc
     1861 caaggattca gggagcttcc agcatgatgt ggctgcaaag cccgagaaag gcttcagaac
     1921 agcacagaac ctagaggcag aggcgtcccc tgaggaagaa ctcccagaag cagatgacct
     1981 ggatggactc ctgagtgagc ttcctgaaga cttcttctgt gggaccagta gtggtcatca
     2041 ccatcaccat cactaactcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     2101 gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca
     2161 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     2221 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2281 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc
     2341 gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2401 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     2461 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2521 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2581 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2641 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2701 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2761 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2821 acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2881 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2941 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3001 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3061 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3121 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3181 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     3241 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     3301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcag
     3361 cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     3421 gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga taaagccgga
     3481 aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ttggtaactg
     3901 atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa agaacagatt
     3961 ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta gctcagcgaa
     4021 ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat gatgaatcaa
     4081 tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg caatttatct
     4141 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     4201 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4261 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4381 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     4441 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     4501 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     4561 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     4621 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     4681 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     4741 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     4801 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     4861 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     4921 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     4981 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     5041 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     5101 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     5161 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     5221 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     5281 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     5341 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     5401 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     5461 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaa
//
