LOCUS       sequence-75424-e        5896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. Coli N-term HexHis Tagged Protein Expression Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  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 5896)
  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
FEATURES             Location/Qualifiers
     source          1..5896
                     /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(2079..2096)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2249..2335
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2427..2454
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2474..2565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2566..2634,2635..3426)
                     /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(2784..2803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3597..4185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4086..4105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4339..4356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4371..4511
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4437..4456)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4597..4619
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4697..4774
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4775..5857
                     /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(4794..4813)
                     /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 cgcgcgatcg
      541 atcgggagga ggcgggagta gcgacggcag cgggcgggca gctggcaggc gggcgtcccg
      601 cagtagcggg cgggcccggc gggggcgcca cgagccgggg ctggggggcc cggcggagcg
      661 cggcgcgggg gaggcacggc tggaagaggc agtcaatcgc tgggtgctca agttctactt
      721 ccacgaggcg ctgcgggcct ttcggggtag ccggtacggg gacttcagac agatccggga
      781 catcatgcag gctttgcttg tcaggccctt ggggaaggag cacaccgtgt cccgattgct
      841 gcgggttatg cagtgtctgt cgcggattga agaaggggaa aatttagact gttcctttga
      901 tatggaggct gagctcacac cactggaatc agctatcaat gtgctggaga tgattaaaac
      961 ggaatttaca ctgacagaag cagtggtcga atccagtaga aaactggtca aggaagctgc
     1021 tgtcattatt tgtatcaaaa acaaagaatt tgaaaaggct tcaaaaattt tgaaaaaaca
     1081 tatgtccaag gaccccacaa ctcagaagct gagaaatgat ctcctgaata ttattcgaga
     1141 aaagaacttg gcccatcctg ttatccagaa cttttcatat gagaccttcc agcagaagat
     1201 gctgcgcttc ctggagagcc acctggatga cgccgagccc tacctcctca cgatggccaa
     1261 aaaggctttg aaatctgagt ccgctgcctc aagtacaggg aaggaagata aacagccagc
     1321 accagggcct gtggaaaagc cacccagaga acccgcaagg cagctacgga atcctccaac
     1381 caccattgga atgatgactc tgaaagcagc tttcaagact ctgtctggtg cacaggattc
     1441 tgaggcagcc tttgcaaaac tggaccagaa ggatctggtt cttcctactc aagctctccc
     1501 agcatcacca gccctcaaaa acaagagacc cagaaaagat gaaaacgaaa gttcagcccc
     1561 ggctgacggt gagggtggct cggaactgca gcccaagaac aagcgcatga caataagcag
     1621 attggtcttg gaggaggaca gccagagtac tgagcccagc gcaggcctca actcctccca
     1681 ggaggccgct tcagcgccac catccaagcc caccgttctc aaccaacccc tccctggaga
     1741 gaagaatccc aaagtaccca aaggcaagtg gaacagctct aatggggttg aagaaaagga
     1801 gacttgggtg gaagaggatg aactgtttca agttcaggca gcaccagatg aagacagtac
     1861 aaccaatata acaaaaaagc agaagtggac tgtagaagaa agcgagtggg tcaaggctgg
     1921 agtgcagaaa tatggggaag gaaactgggc tgccatttct aaaaattacc catttgttaa
     1981 ccgaacagct gtgatgatta aggatcgctg gcggaccatg aaaagacttg gcatgaactg
     2041 aaagcttggc tgtttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     2101 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     2161 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     2221 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2281 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     2341 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     2401 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     2461 tctacaaact ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2521 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2581 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2641 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2701 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2761 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     2821 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2881 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2941 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3001 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3061 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3121 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3181 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3241 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3301 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3361 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3421 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3481 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3541 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3601 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3661 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3721 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3781 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3841 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3901 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3961 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4021 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4081 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4141 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4201 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4261 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4321 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4381 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     4441 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     4501 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4561 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4621 tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc
     4681 ggcatgcatt tacgttgaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     4741 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     4801 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     4861 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     4921 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     4981 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     5041 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     5101 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     5161 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     5221 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     5281 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     5341 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     5401 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     5461 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     5521 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     5581 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat
     5641 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     5701 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     5761 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5821 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5881 agcgcgaatt gatctg
//
