LOCUS       sequence-376964-        8278 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 8278)
  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 8278)
  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: PABPC1
FEATURES             Location/Qualifiers
     source          1..8278
                     /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(3235..3259)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3384..3425
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3880..3897)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3886..4110
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4243..4262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4394..4441
                     /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             4449..5471
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4449..5471
                     /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    5601..5722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5638..5657)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5692..5711
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5771..5787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5771..5787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5784..5806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5795..5811
                     /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(5819..5825,5826..5843,5844..5849))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5864..5885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6002..6019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6173..6761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6253..6272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6932..7723,7724..7792))
                     /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     7555..7574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7793..7897)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7972..7991)
                     /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 ccatgaaccc cagtgccccc agctacccca tggcctcgct ctacgtgggg gacctccacc
     1381 ccgacgtgac cgaggcgatg ctctacgaga agttcagccc ggccgggccc atcctctcca
     1441 tccgggtctg cagggacatg atcacccgcc gctccttggg ctacgcgtat gtgaacttcc
     1501 agcagccggc ggacgcggag cgtgctttgg acaccatgaa ttttgatgtt ataaagggca
     1561 agccagtacg catcatgtgg tctcagcgtg atccatcact tcgcaaaagt ggagtaggca
     1621 acatattcat taaaaatctg gacaaatcca ttgataataa agcactgtat gatacatttt
     1681 ctgcttttgg taacatcctt tcatgtaagg tggtttgtga tgaaaatggt tccaagggct
     1741 atggatttgt acactttgag acgcaggaag cagctgaaag agctattgaa aaaatgaatg
     1801 gaatgctcct aaatgatcgc aaagtatttg ttggacgatt taagtctcgt aaagaacgag
     1861 aagctgaact tggagctagg gcaaaagaat tcaccaatgt ttacatcaag aattttggag
     1921 aagacatgga tgatgagcgc cttaaggatc tctttggcaa gtttgggcct gccttaagtg
     1981 tgaaagtaat gactgatgaa agtggaaaat ccaaaggatt tggatttgta agctttgaaa
     2041 ggcatgaaga tgcacagaaa gctgtggatg agatgaacgg aaaggagctc aatggaaaac
     2101 aaatttatgt tggtcgagct cagaaaaagg tggaacggca gacggaactt aagcgcaaat
     2161 ttgaacagat gaaacaagat aggatcacca gataccaggg tgttaatctt tatgtgaaaa
     2221 atcttgatga tggtattgat gatgaacgtc tccggaaaga gttttctcca tttggtacaa
     2281 tcactagtgc aaaggttatg atggagggtg gtcgcagcaa agggtttggt tttgtatgtt
     2341 tctcctcccc agaagaagcc actaaagcag ttacagaaat gaacggtaga attgtggcca
     2401 caaagccatt gtatgtagct ttagctcagc gcaaagaaga gcgccaggct cacctcacta
     2461 accagtatat gcagagaatg gcaagtgtac gagctgttcc caaccctgta atcaacccct
     2521 accagccagc acctccttca ggttacttca tggcagctat cccacagact cagaaccgtg
     2581 ctgcatacta tcctcctagc caaattgctc aactaagacc aagtcctcgc tggactgctc
     2641 agggtgccag acctcatcca ttccaaaata tgcccggtgc tatccgccca gctgctccta
     2701 gaccaccatt tagtactatg agaccagctt cttcacaggt tccacgagtc atgtcaacac
     2761 agcgtgttgc taacacatca acacagacaa tgggtccacg tcctgcagct gcagccgctg
     2821 cagctactcc tgctgtccgc accgttccac agtataaata tgctgcagga gttcgcaatc
     2881 ctcagcaaca tcttaatgca cagccacaag ttacaatgca acagcctgct gttcatgtac
     2941 aaggtcagga acctttgact gcttccatgt tggcatctgc ccctcctcaa gagcaaaagc
     3001 aaatgttggg tgaacggctg tttcctctta ttcaagccat gcaccctact cttgctggta
     3061 aaatcactgg catgttgttg gagattgata attcagaact tcttcatatg ctcgagtctc
     3121 cagagtcact ccgttctaag gttgatgaag ctgtagctgt actacaagcc caccaagcta
     3181 aagaggctgc ccagaaagca gttaacagtg ccaccggtgt tccaactgtt ttggacccag
     3241 ctttcttgta caaagtggtt gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     3301 cgcggttcga aaccagcgga ggcggcggaa gcgggggagg aggcagcggc ggaggcggca
     3361 gcgggggcgg cggaagcgct accggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3421 ctacgcgtac cggtatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa
     3481 ctggcgacgt gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct
     3541 ctaactcgcg atcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga
     3601 atcgcaaata caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg
     3661 aagagcttcc tgtagccgga tggagatctt acttaaatat ggaactaacc attccaattt
     3721 tcgccacgaa ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg
     3781 gaaacccgat tccctcggcc atcgcggcca actccggcat ctacggaggt ggaggtggat
     3841 ctcgtacggg ttagtaatga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3901 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3961 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4021 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4081 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     4141 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4201 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4261 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4321 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4381 gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
     4441 ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4501 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4561 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4621 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     4681 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4741 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     4801 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4861 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4921 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4981 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     5041 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5101 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5161 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5221 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5281 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5341 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5401 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5461 caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5521 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5581 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5641 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5701 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5761 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5821 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5881 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5941 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7921 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     7981 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8041 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     8101 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     8161 ttgacattga ttattgacta ggcttttgca aaaagctttg caaagatgga taaagtttta
     8221 aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaatt
//
