LOCUS       sequence-235190-        6667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for wild-type Bxb1 with a C-terminal 
            His-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6667)
  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 6667)
  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-Bxb1-His
FEATURES             Location/Qualifiers
     source          1..6667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             71..1570
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     CDS             1580..1597
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1613..1657
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1695..1713)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1709..1756
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1793..2248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1880..1899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2090..2111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2363..3154,3155..3223))
                     /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     2986..3005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3224..3316)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3397..3985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3886..3905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4139..4156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4171..4313
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4239..4258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4399..4421
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4415..4606)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5178..6260)
                     /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     6222..6241
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6261..6338)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6544..6563
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6651..6667,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6651..6667,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgagagccc tggtagtcat ccgcctgtcc cgcgtcaccg atgctacgac
      121 ttcaccggag cgtcagctgg agtcttgcca gcagctctgc gcccagcgcg gctgggacgt
      181 cgtcggggta gcggaggatc tggacgtctc cggggcggtc gatccgttcg accggaagcg
      241 cagaccgaac ctggcccggt ggctagcgtt cgaggagcaa ccgttcgacg tgatcgtggc
      301 gtaccgggta gaccggttga cccgatcgat ccggcatctg cagcagctgg tccactgggc
      361 cgaggaccac aagaagctgg tcgtctccgc gaccgaagcg cacttcgata cgacgacgcc
      421 gtttgcggcg gtcgtcatcg cgcttatggg aacggtggcg cagatggaat tagaagcgat
      481 caaagagcgg aaccgttcgg ctgcgcattt caatatccgc gccgggaaat accgaggatc
      541 cctgccgccg tggggatacc tgcctacgcg cgtggacggg gagtggcggc tggtgccgga
      601 ccctgtgcag cgagagcgca tcctcgaggt gtatcaccgc gtcgtcgaca accacgagcc
      661 gctgcacctg gtggcccacg acctgaaccg gcgtggtgtc ctgtcgccga aggactactt
      721 cgcgcagctg caaggccgcg agccgcaggg ccgggagtgg tcggctaccg cgctgaagcg
      781 atcgatgatc tccgaggcga tgctcgggta cgcgactctg aacggtaaga ccgtccgaga
      841 cgacgacgga gccccgctgg tgcgggctga gccgatcctg acccgtgagc agctggaggc
      901 gctgcgcgcc gagctcgtga agacctcccg ggcgaagccc gcggtgtcta ccccgtcgct
      961 gctgctgcgg gtgttgttct gcgcggtgtg cggggagccc gcgtacaagt tcgccggggg
     1021 aggacgtaag cacccgcgct accgctgccg ctcgatgggg ttcccgaagc actgcgggaa
     1081 cggcacggtg gcgatggccg agtgggacgc gttctgcgag gagcaggtgc tggatctgct
     1141 cggggacgcg gagcgtctgg agaaagtctg ggtagccggc tcggactccg cggtcgaact
     1201 cgcggaggtg aacgcggagc tggtggacct gacgtcgctg atcggctccc cggcctaccg
     1261 ggccggctct ccgcagcgag aagcactgga tgcccgtatt gcggcgctgg ccgcgcggca
     1321 agaggagctg gagggtctag aggctcgccc gtctggctgg gagtggcgcg agaccgggca
     1381 gcggttcggg gactggtggc gggagcagga caccgcggca aagaacacct ggcttcggtc
     1441 gatgaacgtt cggctgacgt tcgacgtccg cggcgggctg actcgcacga tcgacttcgg
     1501 ggatctgcag gagtacgagc agcatctcag gctcggcagc gtggtcgaac ggctacacac
     1561 cgggatgtcg ggcggatccc atcaccacca ccatcactga ctcgagtctg gtaaagaaac
     1621 cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac
     1681 ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1741 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     1801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1861 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1981 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2101 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2161 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2221 tttaacaaaa tattaacgtt tacaatttct ggcggcacga tggcatgaga ttatcaaaaa
     2281 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2341 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2401 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2461 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2521 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2581 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2641 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2701 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2761 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2821 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2881 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2941 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3001 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3061 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3121 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3181 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3241 catgattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3301 tagaaaaata aacaaatagg tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3541 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4081 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4141 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4201 gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca
     4261 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     4321 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4381 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4441 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     4501 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     4561 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     4621 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     4681 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     4741 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     4801 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4861 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     4921 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     4981 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     5041 tcaacgacag gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg
     5101 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5281 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5341 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     5401 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     5461 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     5521 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     5581 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     5641 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     5701 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     5761 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     5821 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     5881 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     5941 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6001 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6061 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6121 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6181 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6241 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6301 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6361 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     6421 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     6481 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     6541 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     6601 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     6661 cactata
//
