LOCUS       sequence-238573-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for Tre with an N-terminal His-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  AUTHORS   Bessen JL, Afeyan LK, Dancik V, Koblan LW, Thompson DB, Leichner C, 
            Clemons PA, Liu DR
  TITLE     High-resolution specificity profiling and off-target prediction for 
            site-specific DNA recombinases.
  JOURNAL   Nat Commun. 2019 Apr 26;10(1):1937. doi: 10.1038/s41467-019-09987-0.
  PUBMED    31028261
REFERENCE   2  (bases 1 to 6196)
  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"; journalName: "Nat 
            Commun."; date: "2019-04-26"; volume: "10(1):1937. doi"; pages: " 
            10.1038/s41467-019-09987-0"
COMMENT     Sequence Label: pET-His-Tre
FEATURES             Location/Qualifiers
     source          1..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    112..136
                     /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)."
     RBS             151..173
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             186..203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1254..1298
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1336..1354)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1350..1397
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1434..1889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1521..1540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1731..1752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2004..2795,2796..2864))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2627..2646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2865..2957)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3038..3626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3527..3546
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3780..3797
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3812..3951
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3877..3896)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4037..4059
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4053..4244)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4816..5898)
                     /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     5860..5879
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5899..5976)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6182..6196,1..5)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
       61 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
      121 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca
      181 tggcgcatca ccaccaccat cacggcggga gcagcaattt actgaccctg caccatagtt
      241 tgcctgcatt accggccgat gcaacgagtg atgaggttcg caagaacctg atggacgtgt
      301 tcagggatcg cccggcgttt tctgagcata cctgggaaat gcttctgtcc gtttgccggt
      361 cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg
      421 ttcgcgatta tcttctacat cttcaggcgc gcggtctggc agtaaaaact atccagcaac
      481 atttgtgcag gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca
      541 atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgta
      601 ccaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa
      661 atagcgatcg ctgccaggat atacgtaatc tggcatttct gggggttgct tataacaccc
      721 tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg
      781 ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga
      841 aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag
      901 ctgatgatcc gaataactac ctgttttgcc gggtcagacg ttatggtgtt gccgcgccat
      961 ctgccaccag ccagctatca acttacgccc tgcagcgcat ttttgaagca actcatcgat
     1021 tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg
     1081 cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc
     1141 aagctggtgg ctggaccacc gtaaatagcg tcatgaacta tatccgtaac ctggatagtg
     1201 aaacaggggc aatggtgcgc ctgctggaag atggcgacta actcgagtct ggtaaagaaa
     1261 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1321 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1381 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     1441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1621 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1741 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1801 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1861 ttttaacaaa atattaacgt ttacaatttc tggcggcacg atggcatgag attatcaaaa
     1921 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1981 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2041 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2101 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2161 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2221 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2281 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2341 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2401 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2461 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2521 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2581 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2641 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2701 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2761 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2821 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2881 tcatgattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2941 ttagaaaaat aaacaaatag gtcatgacca aaatccctta acgtgagttt tcgttccact
     3001 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3061 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3121 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3181 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3241 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3301 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3361 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3421 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3481 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3541 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3601 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3661 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3721 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3781 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3841 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3901 gttaagccag tatacactca gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3961 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4021 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4081 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4141 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4201 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4261 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4321 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4381 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4441 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4501 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4561 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4621 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4681 aacgacagga gcacgatcat gctagtcatg ccccgcgccc accggaagga gctgactggg
     4741 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4801 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     4921 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     4981 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5041 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatccaacta ccgagatgtc
     5101 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5161 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5221 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5281 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5341 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5401 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5461 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5521 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5581 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5641 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5701 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5761 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5821 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     5881 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     5941 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6001 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6061 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6121 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccacat
     6181 cggtgatgtc ggcgat
//
