LOCUS       sequence-376021-        7051 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 7051)
  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 7051)
  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: CPSF5
FEATURES             Location/Qualifiers
     source          1..7051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..1298
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          351..1289
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1246..1266
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1315..1334
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1315..1333
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1412..1436
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2127..2151)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2276..2317
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2772..2789)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2778..3002
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3135..3154)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3286..3333
                     /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             3341..4363
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3341..4363
                     /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    4493..4614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4530..4549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4584..4603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4663..4679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4663..4679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4676..4698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4687..4703
                     /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(4711..4717,4718..4735,4736..4741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4756..4777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4894..4911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5065..5653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5145..5164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5824..6615,6616..6684))
                     /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     6447..6466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6685..6789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6864..6883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttgacattg attattgact aggcttttgc aaaaagcttt gcaaagatgg ataaagtttt
       61 aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg
      121 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
      181 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      241 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      301 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      361 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      421 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
      481 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
      541 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
      601 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
      661 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
      721 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
      781 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
      841 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
      901 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
      961 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     1021 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     1081 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     1141 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     1201 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     1261 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactaatac
     1321 gactcactat agggagaccc aagctggcta ggtaagcttg gtaccgagct cggatccact
     1381 agtccagtgt ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggctccac
     1441 catgtctgtg gtaccgccca atcgctcgca gaccggctgg ccccgggggg tcactcagtt
     1501 cggcaacaag tacatccagc agacgaagcc cctcaccctg gagcgcacca tcaacctgta
     1561 ccctcttacc aattatactt ttggtacaaa agagcccctc tacgagaagg acagctctgt
     1621 tgcagccaga tttcagcgca tgagggaaga atttgataaa attggaatga ggaggactgt
     1681 agaaggggtt ctgattgtac atgagcaccg gctaccccat gtgttactgc tgcagctggg
     1741 aacaactttc ttcaaactac ctggtggtga acttaaccca ggagaagatg aagttgaagg
     1801 actaaaacgc ttaatgacag agatactggg tcgtcaggat ggagttttgc aagactgggt
     1861 cattgacgat tgcattggta actggtggag accaaatttt gaacctcctc agtatccata
     1921 tattcctgca catattacaa agcctaagga acataagaag ttgtttctgg ttcagcttca
     1981 agaaaaagcc ttgtttgcag tccctaaaaa ttacaagctg gtagctgcac cattgtttga
     2041 attgtatgac aatgcaccag gatatggacc catcatttct agtctccctc agctgttgag
     2101 caggttcaat tttatttaca acttggaccc agctttcttg tacaaagtgg ttgatatcca
     2161 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaaccagcg gaggcggcgg
     2221 aagcggggga ggaggcagcg gcggaggcgg cagcgggggc ggcggaagcg ctaccggtaa
     2281 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtatgg cttctaactt
     2341 tactcagttc gttctcgtcg acaatggcgg aactggcgac gtgactgtcg ccccaagcaa
     2401 cttcgctaac ggggtcgctg aatggatcag ctctaactcg cgatcacagg cttacaaagt
     2461 aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt
     2521 gcctaaagtg gcaacccaga ctgttggtgg tgaagagctt cctgtagccg gatggagatc
     2581 ttacttaaat atggaactaa ccattccaat tttcgccacg aattccgact gcgagcttat
     2641 tgttaaggca atgcaaggtc tcctaaaaga tggaaacccg attccctcgg ccatcgcggc
     2701 caactccggc atctacggag gtggaggtgg atctcgtacg ggttagtaat gagtttaaac
     2761 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2821 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2881 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2941 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3001 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3061 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3121 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3181 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3241 cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt tcctattccg
     3301 aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg aactcaccgc
     3361 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     3421 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     3481 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     3541 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     3601 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     3661 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     3721 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     3781 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     3841 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     3901 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     3961 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4021 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     4081 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     4141 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     4201 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     4261 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     4321 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt
     4381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4501 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4561 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4621 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6781 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6841 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6901 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6961 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     7021 ctgcttcgcg atgtacgggc cagatatacg c
//
