LOCUS       sequence-272888-        7191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBP-tagged ORF24-NTD 3L_A 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-3L_A
FEATURES             Location/Qualifiers
     source          1..7191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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     13..35
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             94..1194
                     /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     1168..1191
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     complement(1848..1865)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1848..1864)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1857..1879)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2043..2060)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2213..2299
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2391..2418
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2437..2528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2529..2597,2598..3389)
                     /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(2747..2766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3431..3944)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3626..3647)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3838..3857
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4055..4643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4544..4563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4797..4814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4829..4971
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4897..4916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5057..5079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5073..5264)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5760..5837
                     /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             5838..6920
                     /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(5857..5876)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7163..7168,7169..7184,7185..7191)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
       61 tcacgagcac ttcaccaaca aggaccatag catatgaaaa tcgaagaagg taaactggta
      121 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
      181 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
      241 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
      301 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
      361 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
      421 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
      481 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
      541 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
      601 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
      661 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
      721 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
      781 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
      841 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
      901 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
      961 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     1021 gaagagttgg cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa
     1081 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     1141 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg
     1201 agctcaatgg cagcgctcga gggcccccta ctactgccac cgagcgcctc cctgacgacg
     1261 agtccgcaga ccacgtgtta tcaagcgact tgggaatcac agctggaaat attctgctgt
     1321 ctggccacca actcgcacct gcaggcagag ctgaccttag aaggtcttga taagatgatg
     1381 cagcccgagc ccaccttttt cgcctgcaga gcgatacgca gagcagccgc gggggaacgc
     1441 ctccaccctt ttatacatca agaagggact cttttgggaa aagtgggtcg acggtacagc
     1501 ggcgaaggtt taataattga cggtggtgga gtgtttacgc gcggacagat agacaccgac
     1561 aactacctac ctgcggtggg atcatgggaa cttaccgatg attgtgataa accctgcgaa
     1621 ttcagggagc tacgctcgct gtatcttccc gcgctactaa cgtgcaccat atgttacaaa
     1681 gccatgttca ggatagtgtg caggtacctg gagttctggg agttcgaaca gtgttttcat
     1741 gcgtttctgg cggtgttgcc ccatagtcta caacccacaa tctatcaaaa ttattttgca
     1801 ctcctggagt aaggatcctc tagagtcgac ctgcaggcaa gcttggcact ggccgtcgtt
     1861 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     1921 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1981 ttgcgcagcc tgaatggcga atggcagctt ggctgttttg gcggatgaga taagattttc
     2041 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     2101 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     2161 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     2221 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2281 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     2341 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     2401 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     2461 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2521 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2581 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2641 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2701 ttcgccccga agaacgtttc ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2761 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2821 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2881 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2941 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3001 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3061 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3121 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3181 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3241 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3301 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3361 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3421 agattgattt accccggttg ataatcagaa aagccccaaa aacaggaaga ttgtataagc
     3481 aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3541 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3601 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3661 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3721 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3781 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3841 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3901 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga
     3961 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4021 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4081 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4141 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4201 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4261 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4321 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4381 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4441 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4501 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4561 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4621 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4681 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4741 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4801 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4861 gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata
     4921 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4981 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5041 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5101 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc acagatgtct
     5161 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     5221 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     5281 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     5341 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     5401 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     5461 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     5521 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     5581 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     5641 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     5701 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgaaattccg
     5761 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     5821 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     5881 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     5941 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     6001 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     6061 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     6121 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     6181 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     6241 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     6301 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     6361 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     6421 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     6481 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     6541 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     6601 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcagtagtg ggatacgacg
     6661 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     6721 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     6781 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     6841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6901 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     6961 caattctcat gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     7021 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     7081 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     7141 gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat g
//
