LOCUS       sequence-458886-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E coli expression vector for Halo-BASU-His biotin ligase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6268)
  AUTHORS   Hanswillemenke A, Hofacker DT, Sorgenfrei M, Fruhner C, 
            Franz-Wachtel M, Schwarzer D, Macek B, Stafforst T
  TITLE     Profiling the interactome of oligonucleotide drugs by proximity 
            biotinylation.
  JOURNAL   Nat Chem Biol. 2024 Jan 17. doi: 10.1038/s41589-023-01530-z.
  PUBMED    38233583
REFERENCE   2  (bases 1 to 6268)
  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 Chem 
            Biol. 2024 Jan 17. doi: 10.1038/s41589-023-01530-z."
COMMENT     Sequence Label: Halo-BASU-His
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(140..208,209..1000)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(358..377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1171..1759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1660..1679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1929..3011)
                     /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     2973..2992
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3012..3089)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4298..4317
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4298..4316
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4317..4341
                     /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             4356..4378
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4386..5276
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             6096..6113
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6182..6200)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6196..6243
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
       61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      301 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      361 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
      481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
      541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
      601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
      661 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
      721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
      781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
      841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
      901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
      961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1021 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1081 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1801 ctggcctttt gctcatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     1861 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     1921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     2041 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     2101 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     2161 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     2221 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     2281 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     2341 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     2401 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     2461 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     2521 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     2581 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     2641 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     2701 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     2761 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     2821 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     2881 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     2941 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     3001 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     3061 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     3121 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     3181 tggtgcatgc tagatcaatc cgtaaacagg tcaaacatca gttgccgcaa ccaaatattg
     3241 gctaggtcct tgtggtactt cgcatgccag aacatgttga tggctatttc aggcaagacg
     3301 actgggtgcg gcaaggcgct taggccgaag ggctccacgc agcagtcggc taaacgtatc
     3361 ggcacagtgg cgagcagatc ggtgcgctgg aggatgtggc caacggcggc gaagtgcggc
     3421 acttccagac ggatgtcgcg ccggatgccg acccgtgtca tgtacgtgtc cacctcgccg
     3481 tggccggtgc cagcggcgat gacacgcacg tggccgtagg aacagaagcg ctccagagtc
     3541 aggggttcgc gggtgactgg atggtccttg cgacataggc acacgtagtg attctggagc
     3601 agccggcgct gaaagaagcc agtttgcaga ttgggaagca ggcccacggc caagtccacg
     3661 gttccgttct gcaaggcctg catcaggctc atcgaactgt cgcgcaccgt actgatcacg
     3721 caattggggg cctggtgagc cagcacatcc atcagccgcg gcatgaagta gatctcgcca
     3781 atgtcggtca tggccagggt gaaggtacgc tcgctggtca gcggatcgaa gctttcatgg
     3841 tgctgtaggg cgttgcgcag tgcgtgcatg gccgaagtga cgggctcggc cagatgcgcg
     3901 gcatagggtg tgggttccat tccctgatgt gtgcgcacga agagtgggtc ctgtagcgag
     3961 gtgcgcaggc gtttcagcgc attgctcacg gcaggctggg tcaggcccag gttctccgca
     4021 gtgatagaga cgcgtctgtc gaccagcaac tggttgaaca ccaccagcag gtttaaatcc
     4081 aggtcacgca gttccatgtg gcctcgcttg ggttattgct ggtgcccggc cgggcgcaat
     4141 attcatgttg atgatttatt atatatcgag tggtgtattt atcaatattg tttgctccgt
     4201 tatcgttatt aacaagtcat caataaagcc atcacgagta cagcatgaaa aacaataaac
     4261 ttggcttgcg catctaagga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     4321 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     4381 tacatatggc agaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
     4441 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
     4501 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
     4561 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
     4621 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
     4681 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
     4741 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
     4801 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
     4861 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
     4921 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     4981 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     5041 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     5101 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     5161 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     5221 cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt tccggccctg
     5281 caggcggagg cgcgccaggg tctggcggcg gaaaacttag cgagagcgag atccgcttcg
     5341 gcctgaaaac cgaagtgatg ggccagcacc tgatctacca cgacgtgctg agcagcaccc
     5401 agaaaacagc ccacgagctg gccaacaaca atgcccctga gggaacactg gtggtggccg
     5461 ataagcagac agctggccgc ggcggaatga gccgcgtgtg gcacagccaa gaaggcaacg
     5521 gcgtctggat gagcctgatc ctgcgccctg acatccctct ccagaaaacc cctcagctga
     5581 cactgctggc tgctgtggct gtggtgcagg gaattgaaga agccgccgga atccagaccg
     5641 acatcaagtg gcccaacgac atcctgatca acggcaagaa aaccgtgggc atcctgaccg
     5701 agatgcaggc cgaagaggac agagtccgca gcgtgatcat cggcatcggg atcaatgtga
     5761 accagcagcc taacgacttc cccgacgagc tgaaggatat cgccacaagc ctgtctcagg
     5821 ccgctggcga gaagattgat cgcgctggcg tgatccagca catcctgctg tgcttcgaga
     5881 agcggtaccg cgactacatg acccacggct tcacccctat caaactgctg tgggagagct
     5941 acgccctcgg aatcggcacc aacatgcgcg cccgcacact gaacggcacc ttctacggca
     6001 aggccctggg aattgatgac gagggcgttc tgctgctgga aacaaacgag ggcatcaaga
     6061 aaatctacag cgccgacatc agcctgcgcc tcgagcacca ccaccaccac cactaataat
     6121 gactagtcag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     6181 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     6241 ttgctgaaag gaggaactat atccggat
//
