LOCUS       sequence-82839-e        5764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed N-terminally Hexahistine tagged TRF2 Delta Basic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5764)
  AUTHORS   Bower BD, Griffith JD
  TITLE     TRF1 and TRF2 differentially modulate Rad51-mediated telomeric and 
            nontelomeric displacement loop formation in vitro.
  JOURNAL   Biochemistry. 2014 Sep 2;53(34):5485-95. doi: 10.1021/bi5006249. 
            Epub 2014 Aug 21.
  PUBMED    25115914
REFERENCE   2  (bases 1 to 5764)
  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: "10.1021/bi5006249";
            journalName: "Biochemistry"; date: "2014-09-2- 2"; volume: "53"; 
            issue: "34"; pages: "5485-95"
COMMENT     Sequence Label: pTRCHisB-TRF2∆B
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             491..511
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1947..1964)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2117..2203
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2295..2322
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2342..2433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2434..2502,2503..3294)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2652..2671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3465..4053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3954..3973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4207..4224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4239..4379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4305..4324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4465..4487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4565..4642
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4643..5725
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4662..4681)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gaaaacctgt attttcaggg cgatccgagc tcgagatctg cgcgtgatcg
      541 atcgcggctg gaagaggcag tcaatcgctg ggtgctcaag ttctacttcc acgaggcgct
      601 gcgggccttt cggggtagcc ggtacgggga cttcagacag atccgggaca tcatgcaggc
      661 tttgcttgtc aggcccttgg ggaaggagca caccgtgtcc cgattgctgc gggttatgca
      721 gtgtctgtcg cggattgaag aaggggaaaa tttagactgt tcctttgata tggaggctga
      781 gctcacacca ctggaatcag ctatcaatgt gctggagatg attaaaacgg aatttacact
      841 gacagaagca gtggtcgaat ccagtagaaa actggtcaag gaagctgctg tcattatttg
      901 tatcaaaaac aaagaatttg aaaaggcttc aaaaattttg aaaaaacata tgtccaagga
      961 ccccacaact cagaagctga gaaatgatct cctgaatatt attcgagaaa agaacttggc
     1021 ccatcctgtt atccagaact tttcatatga gaccttccag cagaagatgc tgcgcttcct
     1081 ggagagccac ctggatgacg ccgagcccta cctcctcacg atggccaaaa aggctttgaa
     1141 atctgagtcc gctgcctcaa gtacagggaa ggaagataaa cagccagcac cagggcctgt
     1201 ggaaaagcca cccagagaac ccgcaaggca gctacggaat cctccaacca ccattggaat
     1261 gatgactctg aaagcagctt tcaagactct gtctggtgca caggattctg aggcagcctt
     1321 tgcaaaactg gaccagaagg atctggttct tcctactcaa gctctcccag catcaccagc
     1381 cctcaaaaac aagagaccca gaaaagatga aaacgaaagt tcagccccgg ctgacggtga
     1441 gggtggctcg gaactgcagc ccaagaacaa gcgcatgaca ataagcagat tggtcttgga
     1501 ggaggacagc cagagtactg agcccagcgc aggcctcaac tcctcccagg aggccgcttc
     1561 agcgccacca tccaagccca ccgttctcaa ccaacccctc cctggagaga agaatcccaa
     1621 agtacccaaa ggcaagtgga acagctctaa tggggttgaa gaaaaggaga cttgggtgga
     1681 agaggatgaa ctgtttcaag ttcaggcagc accagatgaa gacagtacaa ccaatataac
     1741 aaaaaagcag aagtggactg tagaagaaag cgagtgggtc aaggctggag tgcagaaata
     1801 tggggaagga aactgggctg ccatttctaa aaattaccca tttgttaacc gaacagctgt
     1861 gatgattaag gatcgctggc ggaccatgaa aagacttggc atgaactgaa agcttggctg
     1921 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1981 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2041 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2101 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2161 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2221 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2281 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     2341 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2401 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2461 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2521 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2581 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2641 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     2701 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2761 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2821 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2881 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2941 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3001 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3061 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3121 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3181 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3241 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3301 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3361 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3421 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3541 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3601 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3661 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3721 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3781 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3841 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3901 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3961 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4021 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4081 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4141 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4201 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4261 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     4321 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     4381 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4441 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4501 caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta
     4561 cgttgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     4621 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     4681 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     4741 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     4801 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     4861 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     4921 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     4981 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     5041 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     5101 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     5161 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     5221 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     5281 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     5341 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     5401 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     5461 cgacgatacc gaagacagct catgttatat cccgccgtca accaccatca aacaggattt
     5521 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     5581 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     5641 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5701 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag cgcgaattga
     5761 tctg
//
