LOCUS       sequence-376047-        8107 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 8107)
  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 8107)
  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: DCP1A
FEATURES             Location/Qualifiers
     source          1..8107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..1198
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          251..1189
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1146..1166
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1215..1234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1215..1233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1312..1336
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3083..3107)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3232..3273
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3728..3745)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3734..3958
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4091..4110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4242..4289
                     /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             4297..5319
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4297..5319
                     /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    5449..5570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5486..5505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5540..5559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5619..5635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5619..5635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5632..5654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5643..5659
                     /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(5667..5673,5674..5691,5692..5697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5712..5733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5850..5867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6021..6609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6101..6120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6780..7571,7572..7640))
                     /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     7403..7422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7641..7745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7820..7839)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
       61 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
      121 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
      181 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
      241 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
      301 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
      361 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
      421 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
      481 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
      541 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
      601 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
      661 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
      721 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
      781 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
      841 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
      901 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
      961 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     1021 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     1081 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     1141 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg
     1201 aattagcttg gtactaatac gactcactat agggagaccc aagctggcta ggtaagcttg
     1261 gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc aacaagtttg
     1321 tacaaaaaag ttggcatgga ggcgctgagt cgagctgggc aggagatgag cctagcggcc
     1381 ctgaagcaac acgaccccta tatcaccagc atcgcagacc tcacgggcca ggtcgctctg
     1441 tacaccttct gccccaaggc caaccagtgg gagaagactg atatagaagg gaccttattc
     1501 gtatatcgaa ggtcagcttc cccttaccat ggttttacca ttgtgaatcg actaaatatg
     1561 cacaatctag ttgaaccagt gaataaagat ttggaatttc agctccatga accatttctt
     1621 ctgtatagaa atgcaagctt gtcgatatat agtatctggt tttatgacaa gaatgactgt
     1681 caccgcatag caaaactcat ggctgatgtg gtagaagagg agacacggcg atcccagcaa
     1741 gctgctcggg acaaacagag tcccagccag gccaatggct gcagcgacca caggcccatc
     1801 gacatcctgg agatgctgag cagagccaag gatgagtatg agaggaatca gatgggtgac
     1861 tcaaatatct ccagccctgg gttacagcca agcactcagc tctccaatct gggaagcacc
     1921 gagactctag aagaaatgcc ctccgggtca caggataagt ctgctccatc tggacacaag
     1981 catctgacgg tagaagagtt atttggaacc tctttgccaa aggaacaacc agcagttgtg
     2041 ggtctggatt cagaagaaat ggagaggttg ccaggagatg cctcccagaa agagcccaat
     2101 tcattcctac catttccctt tgagcagtta ggaggagccc ctcaatcaga aaccctgggt
     2161 gtcccttctg ctgcccacca ttcagtccag cctgaaatca ccaccccggt gctaatcact
     2221 ccagcctcca tcacacagtc caatgaaaag catgctccaa cctacacaat cccgttgagc
     2281 cctgttctca gtcccactct gccagctgaa gctcctactg cacaggttcc ccccagctta
     2341 cctcgaaaca gcaccatgat gcaggcagtg aagaccacgc ctagacagag gtctccactc
     2401 ctgaaccagc cagtccctga gctaagccat gccagtctga ttgccaacca gagccccttc
     2461 agggccccat tgaacgtgac gaacacagct ggcacatccc tcccaagcgt tgatcttctc
     2521 cagaaactca ggttgacccc acagcatgac caaatacaga cacaaccact tgggaaaggt
     2581 gcaatggtag ccagcttttc tccggcagct ggtcagctag ccacacctga gagcttcata
     2641 gagcctccct ctaagacagc agcagcaaga gtggcggcct cagcctccct gagcaacatg
     2701 gtgcttgctc cccttcagtc tatgcagcag aaccaggatc ctgaagtatt tgtgcagcct
     2761 aaggtgttat ccagtgccat cccggttgca ggcgccccac tggttactgc aacgaccact
     2821 gcagtgtctt cagtcctgct ggccccaagt gttttccagc agacagttac aagatcttcg
     2881 gaccttgaga ggaaagccag ctccccttct cctctaacta ttggaacgcc agaaagtcag
     2941 agaaagcctt ccattattct cagcaagtct cagctccagg atacattaat acatctaata
     3001 aagaatgatt ccagcttcct cagtacactt catgaagtct acttgcaggt tctgaccaag
     3061 aacaaagaca accacaacct atgcccaact ttcttgtaca aagtggttga tatccagcac
     3121 agtggcggcc gctcgagtct agagggcccg cggttcgaaa ccagcggagg cggcggaagc
     3181 gggggaggag gcagcggcgg aggcggcagc gggggcggcg gaagcgctac cggtaagcct
     3241 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtatggcttc taactttact
     3301 cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc
     3361 gctaacgggg tcgctgaatg gatcagctct aactcgcgat cacaggctta caaagtaacc
     3421 tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct
     3481 aaagtggcaa cccagactgt tggtggtgaa gagcttcctg tagccggatg gagatcttac
     3541 ttaaatatgg aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt
     3601 aaggcaatgc aaggtctcct aaaagatgga aacccgattc cctcggccat cgcggccaac
     3661 tccggcatct acggaggtgg aggtggatct cgtacgggtt agtaatgagt ttaaacccgc
     3721 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3781 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3841 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3901 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3961 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4021 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4081 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4141 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4201 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     4261 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     4321 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     4381 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     4441 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     4501 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     4561 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4621 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     4681 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4741 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4801 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4861 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4921 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4981 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     5041 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     5101 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     5161 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     5221 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     5281 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     5341 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5401 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5461 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5521 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5581 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5641 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5701 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5761 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5821 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6421 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7081 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7381 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7741 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7801 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     7861 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7921 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     7981 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagg cttttgcaaa
     8041 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     8101 tctaggt
//
