LOCUS       sequence-82846-e        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed N-terminally Hexahistidine tagged TRF1 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  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 5614)
  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-TRF1
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(94..99,100..116,117..123)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    131..147
                     /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     136..158
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    164..233
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             284..310
                     /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             326..343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             347..379
                     /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             392..412
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1797..1814)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1967..2053
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2145..2172
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2192..2283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2284..2352,2353..3144)
                     /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(2502..2521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3315..3903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3804..3823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4057..4074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4089..4229
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4155..4174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4315..4337
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4415..4492
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4493..5575
                     /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(4512..4531)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
       61 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcc ggctcgtata
      121 atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagcgccgc tgagaaaaag
      181 cgaagcggca ctgctcttta acaatttatc agacaatctg tgtgggcact cgaccggaat
      241 tatcgattaa ctttattatt aaaaattaaa gaggtatata ttaatgtatc gattaaataa
      301 ggaggaataa accatggggg gttctcatca tcatcatcat catggtatgg ctagcatgac
      361 tggtggacag caaatgggtc gggatctgta cgaaaacctg tattttcagg gcgatccgag
      421 ctcgagatcc gcgcgtgatc gatcggagga tgtttcctca gcggccccga gcccgcgggg
      481 ctgtgcggat ggtagggatg ccgaccctac tgaggagcag atggcagaaa cagagagaaa
      541 cgacgaggag cagttcgaat gccaggaact gctcgagtgc caggtgcagg tgggggcccc
      601 cgaggaggag gaggaggagg aggaggacgc gggcctggtg gccgaggccg aggccgtggc
      661 tgccggctgg atgctcgatt tcctctgcct ctctctttgc cgagctttcc gcgacggccg
      721 ctccgaggac ttccgcagga cccgcaacag cgcagaggct attattcatg gactatccag
      781 tctaacagct tgccagttga gaacgatata catatgtcag tttttgacaa gaattgcagc
      841 aggaaaaacc cttgatgcac agtttgaaaa tgatgaacga attacaccct tggaatcagc
      901 cctgatgatt tggggttcaa ttgaaaagga acatgacaaa cttcatgaag aaatacagaa
      961 tttaattaaa attcaggcta tagctgtttg tatggaaaat ggcaacttta aagaagcaga
     1021 agaagtcttt gaaagaatat ttggtgatcc aaattctcat atgcctttca aaagcaaatt
     1081 gcttatgata atctctcaga aagatacatt tcattccttt tttcaacact tcagctacaa
     1141 ccacatgatg gagaaaatta agagttatgt gaattatgtg ctaagtgaaa aatcatcaac
     1201 ctttctaatg aaggcagcgg caaaagtagt agaaagcaaa aggacaagaa caataacttc
     1261 tcaagataaa cctagtggta atgatgttga aatggaaact gaagctaatt tggatacaag
     1321 aaaaagtgtt agtgacaaac agtctgcggt aactgaatcc tcagagggta cagtatcctt
     1381 attgaggtct cacaagaatc ttttcttatc taagttgcaa catggaaccc agcaacaaga
     1441 ccttaataag aaagaaagaa gagtaggaac tcctcaaagt acaaaaaaga aaaaagaaag
     1501 cagaagagcc actgaaagca gaatacctgt ttcaaagagt cagccggtaa ctcctgaaaa
     1561 acatcgagct agaaaaagac aggcatggct ttgggaagaa gacaagaatt tgagatctgg
     1621 cgtgaggaaa tatggagagg gaaactggtc taaaatactg ttgcattata aattcaacaa
     1681 ccggacaagt gtcatgttaa aagacagatg gaggaccatg aagaaactaa aactgatttc
     1741 ctcagacagc gaagactgaa agcttggctg tttggcggat gagagaagat tttcagcctg
     1801 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     1861 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     1921 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     1981 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     2041 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     2101 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     2161 ggatggcctt tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat
     2221 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2281 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2341 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2401 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2461 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2521 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2581 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2641 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2701 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2761 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2821 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2881 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2941 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3001 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3061 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3121 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3181 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3241 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3301 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3361 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3421 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3481 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3541 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3601 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3661 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3721 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3781 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3841 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3901 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3961 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4021 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4081 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4141 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     4201 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     4261 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4321 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
     4381 cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt
     4441 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     4501 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4561 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4621 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4681 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     4741 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     4801 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     4861 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     4921 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     4981 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     5041 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     5101 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     5161 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     5221 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     5281 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     5341 cccgccgtca accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5401 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     5461 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5521 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5581 acgcaattaa tgtgagttag cgcgaattga tctg
//
