LOCUS       sequence-338015-        9150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express drosophila dre4 in E.coli expression system. Generated from 
            pDONR-dre4 by Gateway system. Used for custom antibody 
            purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9150)
  AUTHORS   Tettey TT, Gao X, Shao W, Li H, Story BA, Chitsazan AD, Glaser RL, 
            Goode ZH, Seidel CW, Conaway RC, Zeitlinger J, Blanchette M, Conaway
            JW
  TITLE     A Role for FACT in RNA Polymerase II Promoter-Proximal Pausing.
  JOURNAL   Cell Rep. 2019 Jun 25;27(13):3770-3779.e7. doi: 
            10.1016/j.celrep.2019.05.099.
  PUBMED    31242411
REFERENCE   2  (bases 1 to 9150)
  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"; doi: 
            "10.1016/j.celrep.2019.05"; journalName: "Cell Rep"; date: 
            "2019-06-25- 25"; volume: "27"; issue: "13"; pages: "3770-3779.e7"
COMMENT     Sequence Label: pET-DEST42-dre4
FEATURES             Location/Qualifiers
     source          1..9150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     protein_bind    85..109
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3478..3502)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3522..3563
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3573..3590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3647..3665)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3661..3708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3883..3888,3889..3905,3906..3911))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3942..3960)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4028..4132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4133..4201,4202..4993)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4351..4370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5164..5752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5653..5672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5906..5923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5938..6077
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6003..6022)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6163..6185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6179..6370)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7682..8764)
                     /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     8726..8745
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8765..8842)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9048..9067
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggaa tatcacaagt ttgtacaaaa aagcaggctt aatgcgagct
      121 ttgtgctgga caaggaggcg ttcgtgcgcc gcgtgaaacg cctctacatg gaatggaggg
      181 ctcccagcat cggacacgat gacgcgctca ggaatctgga ctgcatcatg agcatcgtgg
      241 gcgtcgaaga ggatgtcatg tactccaagt ccatggccct gcagctctgg ctgctgggct
      301 acgagctgac ggacaccatc tcagtgtttt gctcggatgc cgtgtatttt ctcactagca
      361 aaaagaaaat agaattcctg aagcagacgc agaacatcac cgaagaaggt tttccggaaa
      421 tcaatctgct tgttagggat cggacggaca aggaccaggg caacttcgag aagctcatca
      481 aagccctgca aaactcgaag aagggcaagc gtttgggcgt cttcgccaag gacgcctatc
      541 ccggggagtt cagcgaagca tggaagaaat ccctgacggc gtccaagttc gagcatgtgg
      601 acattagcac tattatcgcc tacctcatgt gtcccaagga tgagtcggag atcaacaaca
      661 tacgcaaggc atcgctagtg tcaatggaca ttttcaacaa gtatttgaaa gatgagatca
      721 tggacattat tgattcggat aggaaagtca aacacaacaa gctctcagat ggctgcgagg
      781 ctgccattgg cgagaagaag tacaccagtg gtctggatcc cagactcctc gatatggcct
      841 acccacccat catccaatcg ggaggagcct acagcctgaa gttctccgcg gtggcggaca
      901 aaaatccctt gcacttcggt gtgattgtgt gctccttggg cgcccgctac aagtcgtatt
      961 gctcaaatat atcgcgcacc tttctggtga atcccaccga ggccatgcag gagaactaca
     1021 catttctggt aagtgtgcag gaggagattc tcaagctgct ggttccgggc acaaagctgt
     1081 gcgacgtcta cgaaaagacc ctggacttcg tgaagaagga gaagccgagt atggtggaca
     1141 atctgcccaa aagctttggt ttcgccatgg gcctagagtt ccgggagaac tccattgtca
     1201 tcggacccaa atgccaggcg ctactcaaga agaatatggt ctttaatttg cacgtgggca
     1261 tctcgaatct gaccaatccc gaagccaccg acaaagaggg caagaactac gccctcttca
     1321 tcggagacac tgtgctagta ggcgaacaat ccccagccag tgtgatgact ccgtccaaga
     1381 agaagatcaa gaacgtcggc atctttatca aggacgacag cgacgaggaa gatgtggacg
     1441 acaagaagac cgctaaagaa gatcagggca ccgagatcct gggccgcagc aagcgcaacg
     1501 cggtgctgga gtcaaagctg cgaaacgaga tcaacacgga ggagaagcgc aaggagcacc
     1561 agcgcgagct ggctcagcag ctgaacgagc gcgccaagga tcggctggcc aggcagggaa
     1621 actcgaagga ggtcgagaaa gtgcgcaaga acaccgtctc ctacaagtcc atcagccaaa
     1681 tgccgcgcga gccggaagtc aaggagctca agctttacgt ggataagaag tacgagaccg
     1741 tcatcatgcc agtgttcggc atccaggtgc cgttccatat atccaccatc aagaacatct
     1801 cgcagtcggt ggagggcgag tacacgtact tgcgtatcaa cttctttcat cccggcgcca
     1861 caatgggtcg caacgaaggc ggactctatc ctcagccaga ggccacattt gtcaaggaag
     1921 ttacataccg cagctctaat gttaaggaac acggggaggt gggagctcca tccgcaaatc
     1981 taaacaatgc tttccggctg atcaaggagg tgcagaagcg cttcaagacc cgcgaggctg
     2041 aggagcgcga aaaggaggat cttgtgaagc aagacacact cattctgtcc cagaacaagg
     2101 gtaatcccaa gctgaaggac ctatacattc ggcccaacat cgtgaccaag cgcatgacgg
     2161 gcagcctgga agcgcacagc aatggattcc gctacatctc cgtgcgcggc gacaaggtgg
     2221 acatcttgta caacaacatc aaaagcgcgt ttttccagcc ctgcgatggc gagatgatca
     2281 ttcttctgca tttccacctg aagtacgcca ttatgtttgg caaaaagaag cacgtggatg
     2341 tacagttcta cacggaagtg ggagagatca ccacggatct gggcaagcac cagcacatgc
     2401 acgaccgcga cgacttggct gctgagcagg cagaacgtga gctgcggcac aagctcaaga
     2461 ctgccttcaa gagcttctgc gagaaggtgg aaaccatgac caagtcggta gtggagttcg
     2521 acacgccgtt ccgtgagttg ggattccctg gagcaccctt ccgcagcacg gtgacgctgc
     2581 aacccacctc gggctccctg gtcaatctga cggaatggcc accatttgtc atcacgctgg
     2641 acgatgtgga gttggtgcac ttcgagcgcg ttcagttcca cttgcgcaac ttcgatatga
     2701 tatttgtgtt caaggagtac aataagaagg ttgccatggt gaacgccatt cctatgaaca
     2761 tgctggatca cgtgaaggag tggctcaact cctgcgacat ccgctattcg gagggtgttc
     2821 aatccctgaa ctggcaaaag atcatgaaga ccatcaccga cgatcctgag ggattctttg
     2881 aacaaggcgg ctggacattc ttggacccgg aatcgggcag cgaaggcgaa aacgaaaccg
     2941 ccgaatccga agaggacgag gcctacaatc ccaccgacgc cgaatcggat gaggagtcgg
     3001 acgaagattc cgagtactca gaggcatcag aagacagcga ggaaagcgat gaggaccttg
     3061 gctctgatga ggaatctggc aaggattggt ccgatctgga gcgcgaggct gctgaggaag
     3121 accgcaacca cgattatgcc gcagatgaca aaccacgcaa cggcaaattc gattcgaaga
     3181 agcacggcaa gagttcaaag cacagtcgcc gcgaccgcga ggaggctcga tcgtcgtcgc
     3241 acagtaaaaa gcacaagtcg aactcctcct cgtcctcttc gcatttaaag tcctccagct
     3301 ccaaacatgg cagctcatct agcccgtcga agtcgtcgaa ggacaagtac aacagccggg
     3361 acaagcatca ttcctcctca tcgtccggaa acaagagcag cagcaaggac aaggatcgca
     3421 agcgctcgcg ggatgacagt cgcgacaatg ggcacaagtc gaagaagtcg cgacattacc
     3481 cagctttctt gtacaaagtg gtgatcaatt cgaagcttga aggtaagcct atccctaacc
     3541 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttgat
     3601 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3661 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     3721 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     3781 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     3841 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     3901 taagctgtca aacatgagaa ttaattcttg aagacgaaag ggcctcgtga tacgcctatt
     3961 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5101 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5161 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5281 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5821 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5881 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     5941 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     6001 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     6061 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6121 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6181 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     6241 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     6301 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     6361 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     6421 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     6481 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     6541 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     6601 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     6661 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     6721 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     6781 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc
     6841 aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg
     6901 atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc
     6961 caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt
     7021 ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc
     7081 tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat
     7141 cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc
     7201 ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc
     7261 gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg cgaacgccag
     7321 caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa
     7381 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     7441 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     7501 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     7561 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     7621 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     7681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7741 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     7801 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     7861 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     7921 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     7981 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     8041 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     8101 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     8161 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     8221 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     8281 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     8341 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     8401 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     8461 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     8521 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     8581 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     8641 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     8701 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     8761 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     8821 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     8881 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     8941 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     9001 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     9061 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     9121 tagaggatcg agatctcgat cccgcgaaat
//
