LOCUS       sequence-238579-        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"
     CDS             1421..1438
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1457..1501
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1539..1557)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1553..1600
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1637..2092
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1724..1743)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1934..1955
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2207..2998,2999..3067))
                     /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     2830..2849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3068..3160)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3241..3829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3730..3749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3983..4000
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4015..4154
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4080..4099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4240..4262
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4256..4447)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5019..6101)
                     /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     6063..6082
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6102..6179)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6385..6404
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6492..6511
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6492..6510
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6511..6535
                     /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             6550..6572
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(6579..6667,1..1411)
                     /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"
ORIGIN
        1 cgcccagcgc ggctgggacg tcgtcggggt agcggaggat ctggacgtct ccggggcggt
       61 cgatccgttc gaccggaagc gcagaccgaa cctggcccgg tggctagcgt tcgaggagca
      121 accgttcgac gtgatcgtgg cgtaccgggt agaccggttg acccgatcga tccggcatct
      181 gcagcagctg gtccactggg ccgaggacca caagaagctg gtcgtctccg cgaccgaagc
      241 gcacttcgat acgacgacgc cgtttgcggc ggtcgtcatc gcgcttatgg gaacggtggc
      301 gcagatggaa ttagaagcga tcaaagagcg gaaccgttcg gctgcgcatt tcaatatccg
      361 cgccgggaaa taccgaggat ccctgccgcc gtggggatac ctgcctacgc gcgtggacgg
      421 ggagtggcgg ctggtgccgg accctgtgca gcgagagcgc atcctcgagg tgtatcaccg
      481 cgtcgtcgac aaccacgagc cgctgcacct ggtggcccac gacctgaacc ggcgtggtgt
      541 cctgtcgccg aaggactact tcgcgcagct gcaaggccgc gagccgcagg gccgggagtg
      601 gtcggctacc gcgctgaagc gatcgatgat ctccgaggcg atgctcgggt acgcgactct
      661 gaacggtaag accgtccgag acgacgacgg agccccgctg gtgcgggctg agccgatcct
      721 gacccgtgag cagctggagg cgctgcgcgc cgagctcgtg aagacctccc gggcgaagcc
      781 cgcggtgtct accccgtcgc tgctgctgcg ggtgttgttc tgcgcggtgt gcggggagcc
      841 cgcgtacaag ttcgccgggg gaggacgtaa gcacccgcgc taccgctgcc gctcgatggg
      901 gttcccgaag cactgcggga acggcacggt ggcgatggcc gagtgggacg cgttctgcga
      961 ggagcaggtg ctggatctgc tcggggacgc ggagcgtctg gagaaagtct gggtagccgg
     1021 ctcggactcc gcggtcgaac tcgcggaggt gaacgcggag ctggtggacc tgacgtcgct
     1081 gatcggctcc ccggcctacc gggccggctc tccgcagcga gaagcactgg atgcccgtat
     1141 tgcggcgctg gccgcgcggc aagaggagct ggagggtcta gaggctcgcc cgtctggctg
     1201 ggagtggcgc gagaccgggc agcggttcgg ggactggtgg cgggagcagg acaccgcggc
     1261 aaagaacacc tggcttcggt cgatgaacgt tcggctgacg ttcgacgtcc gcggcgggct
     1321 gactcgcacg atcgacttcg gggatctgca ggagtacgag cagcatctca ggctcggcag
     1381 cgtggtcgaa cggctacaca ccgggatgtc gggcggatcc catcaccacc accatcacta
     1441 atgactcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc
     1501 gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata
     1561 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1621 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1681 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1741 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1801 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1861 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1921 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1981 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2041 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttctggcggc
     2101 acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3061 tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta ttgtctcatg
     3121 agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga ccaaaatccc
     3181 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3241 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3301 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3361 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3421 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3481 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3541 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3601 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3661 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3721 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3781 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3841 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3901 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3961 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     4021 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     4081 acaatctgct ctgatgccgc atagttaagc cagtatacac tcagctatcg ctacgtgact
     4141 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4201 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4261 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4321 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4381 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4441 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4501 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4561 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4621 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     4681 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4741 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4801 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4861 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgctagtc atgccccgcg
     4921 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4981 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     5161 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5221 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     5281 tcgtatccaa ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5341 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5401 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5461 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5521 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5581 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5641 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5701 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5761 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5821 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5881 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5941 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6001 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6061 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     6121 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     6181 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     6241 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     6301 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     6361 gtggcgagcc cgatcttcca catcggtgat gtcggcgata taggcgccag caaccgcacc
     6421 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga
     6481 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     6541 agaaataatt ttgtttaact ttaagaagga gatataccat gagagccctg gtagtcatcc
     6601 gcctgtcccg cgtcaccgat gctacgactt caccggagcg tcagctggag tcttgccagc
     6661 agctctg
//
