LOCUS       sequence-165728-        6093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine SRAP domain (250 aa) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Kweon SM, Zhu B, Chen Y, Aravind L, Xu SY, Feldman DE
  TITLE     Erasure of Tet-Oxidized 5-Methylcytosine by a SRAP Nuclease.
  JOURNAL   Cell Rep. 2017 Oct 10;21(2):482-494. doi: 
            10.1016/j.celrep.2017.09.055.
  PUBMED    29020633
REFERENCE   2  (bases 1 to 6093)
  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"; doi: 
            "10.1016/j.celrep.2017.09"; journalName: "Cell Rep"; date: 
            "2017-10-10- 10"; volume: "21"; issue: "2"; pages: "482-494"
COMMENT     Sequence Label: pET30a-SRAP
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(920..964)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(971..988)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(998..1015)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1026..1048
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1063..1087
                     /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)."
     primer_bind     complement(1087..1106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1088..1106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1194..1213)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1419..1496
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1497..2579
                     /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     complement(1516..1535)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3388..3579
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3573..3595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3681..3823
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3736..3755
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3838..3855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4009..4597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4089..4108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4708..4727)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4719..5534
                     /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             4719..5534
                     /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     complement(4792..4811)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5627..6082)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5764..5785)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5976..5995
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtggatgg aaggtgatat
      181 tgtctatggg gtggattagc ttcagagctt cctgagtggc gacctcacca aagtcgagcc
      241 atttggagac tgcttcttct ccatctagta tggcaggcat cctgctgtgg atgtcactca
      301 aacctctgca ggaatccaca gtgatgatgc tgtaggaata caggcactct ccccctggcg
      361 cttcccagca gtcaaagatc cctgccattg tcagcagcct ccagttgtcc cagacctttt
      421 ccttgttgtc agagctgtct gaagcatcgt tcccacctga cttctctgtc ttgatttgag
      481 gaaaatagat gaagtatggt tgcctctggt ttgttccctg acaccgctgc cactcgtaga
      541 atccatctgc taaaacaaca caccgccgtc ctttccccag aggaaccttg aatgactgct
      601 tctccattat ggtatcactg cgacagttgg tagtgttgaa ctgcagctta gaaggatcac
      661 tttctttgaa ccaagatggg actaagcccc atcgcatggg aataattatc cgatctgatg
      721 agtctgcatc cttctcaaag tgcagtctgg agagcagcac ggggctgctg gactgcgggc
      781 tcttgttgta ggaggggcag tacttgtcgg ggtccctcca ctgcgggagc cgccgccggc
      841 cctgccgatc ctgataggcg caggccctgg tgagaacctc tctgggcaag tgacaggacg
      901 ttcgcccgca cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt
      961 ctttcatacc agaaccgcgt ggcaccagac cagaagaatg atgatgatga tggtgcatat
     1021 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
     1081 caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta
     1141 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     1201 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     1261 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     1321 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     1381 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
     1441 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     1501 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     1561 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     1621 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     1681 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     1741 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     1801 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     1861 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     1921 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     1981 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     2041 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     2101 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     2161 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     2221 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     2281 tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     2341 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     2401 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     2461 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     2521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2581 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
     2641 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     2701 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     2761 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     2821 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     2881 ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg
     2941 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     3001 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     3061 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
     3121 ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca
     3181 cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc
     3241 cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca
     3301 tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac
     3361 agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg
     3421 gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg
     3481 gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc
     3541 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3601 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3661 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3721 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg
     3781 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     3841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4441 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
     4681 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4741 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     4801 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     4861 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4921 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4981 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     5041 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     5101 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     5161 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     5221 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     5281 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     5341 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     5401 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     5461 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     5521 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5581 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt
     5641 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5701 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5761 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5821 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5881 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5941 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6001 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6061 aatgcgccgc tacagggcgc gtcccattcg cca
//
