LOCUS       sequence-272729-        7191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBP-tagged ORF24-NTD in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7191)
  AUTHORS   Castaneda AF, Didychuk AL, Louder RK, McCollum CO, Davis ZH, Nogales
            E, Glaunsinger BA
  TITLE     The gammaherpesviral TATA-box-binding protein directly interacts 
            with the CTD of host RNA Pol II to direct late gene transcription.
  JOURNAL   PLoS Pathog. 2020 Sep 4;16(9):e1008843. doi: 
            10.1371/journal.ppat.1008843. eCollection 2020 Sep.
  PUBMED    32886723
REFERENCE   2  (bases 1 to 7191)
  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: "PLoS 
            Pathog."; date: "2020-09-4"; volume: "16(9):e1008843. doi"; pages: "
            10.1371/journal.ppat.1008843. eCollection 2020 Sep"
COMMENT     Sequence Label: pMAL-c2X-ORF24-NTD
FEATURES             Location/Qualifiers
     source          1..7191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(51..56,57..72,73..79)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    87..103
                     /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     92..114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             173..1273
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1247..1270
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     complement(1927..1944)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1927..1943)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1936..1958)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2122..2139)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2292..2378
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2470..2497
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2516..2607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2608..2676,2677..3468)
                     /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(2826..2845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3510..4023)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3917..3936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4134..4722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4623..4642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4876..4893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4908..5050
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4976..4995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5136..5158
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5152..5343)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5839..5916
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5917..6999
                     /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     complement(5936..5955)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
       61 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
      121 ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccatagc atatgaaaat
      181 cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga
      241 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa
      301 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg
      361 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga
      421 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa
      481 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct
      541 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa
      601 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc
      661 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg
      721 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa
      781 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac
      841 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta
      901 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct
      961 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa
     1021 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt
     1081 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga
     1141 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc
     1201 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa
     1261 agacgcgcag actaattcga gctcaatggc agcgctcgag ggccccctac tactgccacc
     1321 gagcgcctcc ctgacgacga gtccgcagac cacgtgttat caagcgactt gggaatcaca
     1381 gctggaaata ttctgctgtc tggccaccaa ctcgcacctg caggcagagc tgaccttaga
     1441 aggtcttgat aagatgatgc agcccgagcc cacctttttc gcctgcagag cgatacgcag
     1501 actactcctg ggggaacgcc tccacccttt tatacatcaa gaagggactc ttttgggaaa
     1561 agtgggtcga cggtacagcg gcgaaggttt aataattgac ggtggtggag tgtttacgcg
     1621 cggacagata gacaccgaca actacctacc tgcggtggga tcatgggaac ttaccgatga
     1681 ttgtgataaa ccctgcgaat tcagggagct acgctcgctg tatcttcccg cgctactaac
     1741 gtgcaccata tgttacaaag ccatgttcag gatagtgtgc aggtacctgg agttctggga
     1801 gttcgaacag tgttttcatg cgtttctggc ggtgttgccc catagtctac aacccacaat
     1861 ctatcaaaat tattttgcac tcctggagta aggatcctct agagtcgacc tgcaggcaag
     1921 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1981 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2041 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcagcttg gctgttttgg
     2101 cggatgagat aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     2161 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     2221 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     2281 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2341 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     2401 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     2461 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     2521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2761 ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa
     2821 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3481 gtttactcat atatacttta gattgattta ccccggttga taatcagaaa agccccaaaa
     3541 acaggaagat tgtataagca aatatttaaa ttgtaaacgt taatattttg ttaaaattcg
     3601 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3661 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3721 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3781 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     3841 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3901 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3961 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4021 cgtaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4081 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4201 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4261 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4321 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4381 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4441 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4501 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4561 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4621 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4681 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4741 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4801 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4861 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4921 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     4981 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     5041 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5101 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5161 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     5221 cagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     5281 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     5341 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     5401 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     5461 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     5521 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     5581 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     5641 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     5701 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     5761 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc
     5821 aacgctgccc gaaattccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga
     5881 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
     5941 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     6001 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     6061 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     6121 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     6181 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     6241 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     6301 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     6361 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     6421 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     6481 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     6541 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     6601 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     6661 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     6721 tcagtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     6781 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     6841 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     6901 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6961 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
     7021 ttagctcact cattaggcac aattctcatg tttgacagct tatcatcgac tgcacggtgc
     7081 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     7141 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg t
//
