LOCUS       sequence-75427-e        5707 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 5707)
  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 5707)
  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∆M
FEATURES             Location/Qualifiers
     source          1..5707
                     /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(1890..1907)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2060..2146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2238..2265
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2285..2376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2377..2445,2446..3237)
                     /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(2595..2614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3408..3996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3897..3916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4150..4167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4182..4322
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4248..4267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4408..4430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4508..4585
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4586..5668
                     /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(4605..4624)
                     /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 gcaccagatt aaaagcttgg
     1861 ctgtttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     1921 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1981 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
     2041 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2101 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     2161 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     2221 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     2281 tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2581 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3541 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4021 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4081 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4141 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4201 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4261 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4321 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4381 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4441 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat
     4501 ttacgttgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
     4561 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     4621 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     4681 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     4741 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     4801 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     4861 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     4921 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     4981 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     5041 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     5101 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     5161 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     5221 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     5281 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     5341 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     5401 atacgacgat accgaagaca gctcatgtta tatcccgccg tcaaccacca tcaaacagga
     5461 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     5521 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     5581 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     5641 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagcgcgaat
     5701 tgatctg
//
