LOCUS       sequence-376755-        8329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For use in RBP tethering screen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8329)
  AUTHORS   Luo EC, Nathanson JL, Tan FE, Schwartz JL, Schmok JC, Shankar A, 
            Markmiller S, Yee BA, Sathe S, Pratt GA, Scaletta DB, Ha Y, Hill DE,
            Aigner S, Yeo GW
  TITLE     Large-scale tethered function assays identify factors that regulate 
            mRNA stability and translation.
  JOURNAL   Nat Struct Mol Biol. 2020 Oct;27(10):989-1000. doi: 
            10.1038/s41594-020-0477-6. Epub 2020 Aug 17.
  PUBMED    32807991
REFERENCE   2  (bases 1 to 8329)
  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: 
            "10.1038/s41594-020-0477-6"; journalName: "Nat Struct Mol Biol"; 
            date: "2020-10"; volume: "27"; issue: "10"; pages: "989-1000"
COMMENT     Sequence Label: FUBP1
FEATURES             Location/Qualifiers
     source          1..8329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3286..3310)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3435..3476
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3931..3948)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3937..4161
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4294..4313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4445..4492
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4500..5522
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4500..5522
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5652..5773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5689..5708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5743..5762
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5822..5838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5822..5838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5835..5857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5846..5862
                     /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)."
     promoter        complement(join(5870..5876,5877..5894,5895..5900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5915..5936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6053..6070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6224..6812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6304..6323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6983..7774,7775..7843))
                     /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     7606..7625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7844..7948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8023..8042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctcca
     1321 ccatggcaga ctattcaaca gtgcctcccc cctcttctgg ctcagctggt ggcggtggtg
     1381 gcggcggtgg tggtggagga gttaacgacg ctttcaaaga tgcactgcag agagcccggc
     1441 agattgcagc aaaaattgga ggtgatgcag ggacatcact gaattcaaat gactatggtt
     1501 atgggggaca aaaaagacct ttagaagatg gagatcaacc agatgctaag aaagttgctc
     1561 ctcaaaatga ctcttttgga acacagttac caccgatgca tcagcagcaa agatctgtaa
     1621 tgacagaaga atacaaagtt ccagatggaa tggttggatt cataattggc agaggaggtg
     1681 aacagatctc acgcatacaa caggaatctg gatgcaaaat acagatagct cctgacagtg
     1741 gtggccttcc agaaaggtcc tgtatgttaa ctggaacacc tgaatctgtc cagtcagcaa
     1801 aacggttact ggaccagatt gttgaaaaag gaagaccagc tcctggcttc catcatggcg
     1861 atggaccggg aaatgcagtt caagaaatca tgattccagc tagcaaggca ggattagtca
     1921 ttggaaaagg gggagaaact attaaacagc ttcaggaacg ggctggagtt aaaatggtta
     1981 tgattcaaga cgggccgcag aacactggtg ctgacaaacc tcttaggatt acaggagacc
     2041 catataaagt tcaacaagcc aaggaaatgg tgttagagtt aattcgtgat caaggcggtt
     2101 tcagagaagt tcggaatgag tatgggtcaa gaataggagg aaatgaaggg atagatgtcc
     2161 ccattccaag atttgctgtt ggcattgtaa taggaagaaa tggagagatg atcaaaaaaa
     2221 tacaaaatga tgctggtgtt cgcattcagt ttaagccaga tgatgggaca acacccgaaa
     2281 ggatagcaca aataacagga cctccagacc gatgtcaaca tgctgcagaa attattacag
     2341 accttcttcg aagtgttcag gctggtaatc ctggtggacc tggacctggt ggtcgaggaa
     2401 gaggtagagg tcaaggcaac tggaacatgg gaccacctgg tggactacag gaatttaatt
     2461 ttattgtgcc aactgggaaa actggattaa taataggaaa aggaggtgaa accataaaaa
     2521 gcataagcca gcagtctggt gcaagaatag aacttcagag aaatcctcca ccaaatgcag
     2581 atcctaatat gaagttattt acaattcgtg gcactccaca acagatagac tatgctcggc
     2641 aactcataga agaaaagatt ggtggcccag taaatccttt agggccacct gtaccccatg
     2701 ggccccatgg tgtcccaggc ccccatggac ctcctgggcc tccagggcct ggaactccaa
     2761 tgggaccata caaccctgca ccttataatc ctggaccacc aggcccggct cctcatggtc
     2821 ctccagcccc atatgctccc cagggatggg gaaatgcata tccacactgg cagcagcagg
     2881 ctcctcctga tccagctaag gcaggaacgg atccaaattc agcagcttgg gctgcttatt
     2941 acgctcacta ttatcaacag caagcacagc caccaccagc agcccctgca ggtgcaccaa
     3001 ctacaactca aactaatgga caaggagatc agcagaatcc agccccagct ggacaggttg
     3061 attataccaa ggcttgggaa gagtactaca agaaaatggg tcaggcagtt cctgctccga
     3121 ctggggctcc tccaggtggt cagccagatt atagtgcagc ctgggctgag tattatagac
     3181 aacaagcagc ctattatgcc cagacaagtc cccagggaat gccacagcat cctccagcac
     3241 ctcagtgcag gtttgatcca gccagtatag aactagctct gttggaccca gctttcttgt
     3301 acaaagtggt tgatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg
     3361 aaaccagcgg aggcggcgga agcgggggag gaggcagcgg cggaggcggc agcgggggcg
     3421 gcggaagcgc taccggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     3481 ccggtatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg
     3541 tgactgtcgc cccaagcaac ttcgctaacg gggtcgctga atggatcagc tctaactcgc
     3601 gatcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat
     3661 acaccatcaa agtcgaggtg cctaaagtgg caacccagac tgttggtggt gaagagcttc
     3721 ctgtagccgg atggagatct tacttaaata tggaactaac cattccaatt ttcgccacga
     3781 attccgactg cgagcttatt gttaaggcaa tgcaaggtct cctaaaagat ggaaacccga
     3841 ttccctcggc catcgcggcc aactccggca tctacggagg tggaggtgga tctcgtacgg
     3901 gttagtaatg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3961 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4021 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4081 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4141 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4201 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4261 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4321 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4381 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4441 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     4501 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     4561 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     4621 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     4681 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     4741 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     4801 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     4861 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     4921 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     4981 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     5041 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     5101 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     5161 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     5221 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     5281 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     5341 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     5401 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     5461 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     5521 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5581 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5641 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5701 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5761 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5821 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5881 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5941 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6001 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6661 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7261 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7981 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     8041 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     8101 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     8161 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     8221 attattgact aggcttttgc aaaaagcttt gcaaagatgg ataaagtttt aaacagagag
     8281 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaatt
//
