LOCUS       sequence-165731-        6090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes murine SRAP domain (250aa) of Hmces/Srap1 with deletion of 
            catalytic Cys2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6090)
  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 6090)
  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-delCys2
FEATURES             Location/Qualifiers
     source          1..6090
                     /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(917..961)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(968..985)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(995..1012)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1023..1045
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1060..1084
                     /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(1084..1103)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1085..1103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1191..1210)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1416..1493
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1494..2576
                     /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(1513..1532)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3385..3576
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3570..3592)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3678..3820
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3733..3752
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3835..3852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4006..4594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4086..4105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4705..4724)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4716..5531
                     /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             4716..5531
                     /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(4789..4808)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5624..6079)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5761..5782)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5973..5992
                     /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 ttcgccccag atctgggctg tccatgtgct ggcgttcgaa tttagcagca gcggtttctt
      961 tcataccaga accgcgtggc accagaccag aagaatgatg atgatgatgg tgcatatgta
     1021 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
     1081 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
     1141 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     1201 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     1261 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     1321 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     1381 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
     1441 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     1501 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     1561 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     1621 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     1681 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     1741 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     1801 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     1861 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     1921 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     1981 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     2041 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     2101 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     2161 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     2221 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     2281 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     2341 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     2401 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     2461 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2521 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2581 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
     2641 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     2701 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     2761 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     2821 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     2881 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
     2941 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     3001 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     3061 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     3121 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     3181 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     3241 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     3301 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     3361 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     3421 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     3481 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     3541 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3601 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3661 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3721 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga
     3781 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     3841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4441 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac
     4681 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
     4741 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     4801 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
     4861 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     4921 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     4981 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     5041 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     5101 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     5161 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     5221 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     5281 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     5341 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     5401 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     5461 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     5521 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat
     5641 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5701 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5761 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5821 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5881 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5941 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6001 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6061 gcgccgctac agggcgcgtc ccattcgcca
//
