LOCUS       sequence-375892-        8464 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 8464)
  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 8464)
  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: ADARB1
FEATURES             Location/Qualifiers
     source          1..8464
                     /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(3421..3445)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3570..3611
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4066..4083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4072..4296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4429..4448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4580..4627
                     /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             4635..5657
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4635..5657
                     /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    5787..5908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5824..5843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5878..5897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5957..5973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5957..5973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5970..5992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5981..5997
                     /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(6005..6011,6012..6029,6030..6035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6050..6071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6188..6205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6359..6947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6439..6458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7118..7909,7910..7978))
                     /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     7741..7760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7979..8083)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8158..8177)
                     /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 gttggcatgg
     1321 atatagaaga tgaagaaaac atgagttcca gcagcactga tgtgaaggaa aaccgcaatc
     1381 tggacaacgt gtcccccaag gatggcagca cacctgggcc tggcgagggc tctcagctct
     1441 ccaatggggg tggtggtggc cccggcagaa agcggcccct ggaggagggc agcaatggcc
     1501 actccaagta ccgcctgaag aaaaggagga aaacaccagg gcccgtcctc cccaagaacg
     1561 ccctgatgca gctgaatgag atcaagcctg gtttgcagta cacactcctg tcccagactg
     1621 ggcccgtgca cgcgcctttg tttgtcatgt ctgtggaggt gaatggccag gtttttgagg
     1681 gctctggtcc cacaaagaaa aaggcaaaac tccatgctgc tgagaaggcc ttgaggtctt
     1741 tcgttcagtt tcctaatgcc tctgaggccc acctggccat ggggaggacc ctgtctgtca
     1801 acacggactt cacatctgac caggccgact tccctgacac gctcttcaat ggttttgaaa
     1861 ctcctgacaa ggcggagcct cccttttacg tgggctccaa tggggatgac tccttcagtt
     1921 ccagcgggga cctcagcttg tctgcttccc cggtgcctgc cagcctagcc cagcctcctc
     1981 tccctgcctt accaccattc ccacccccga gtgggaagaa tcccgtgatg atcttgaacg
     2041 aactgcgccc aggactcaag tatgacttcc tctccgagag cggggagagc catgccaaga
     2101 gcttcgtcat gtctgtggtc gtggatggtc agttctttga aggctcgggg agaaacaaga
     2161 agcttgccaa ggcccgggct gcgcagtctg ccctggccgc catttttaac ttgcacttgg
     2221 atcagacgcc atctcgccag cctattccca gtgagggtct tcagctgcat ttaccgcagg
     2281 ttttagctga cgctgtctca cgcctggtcc tgggtaagtt tggtgacctg accgacaact
     2341 tctcctcccc tcacgctcgc agaaaagtgc tggctggagt cgtcatgaca acaggcacag
     2401 atgttaaaga tgccaaggtg ataagtgttt ctacaggaac aaaatgtatt aatggtgaat
     2461 acatgagtga tcgtggcctt gcattaaatg actgccatgc agaaataata tctcggagat
     2521 ccttgctcag atttctttat acacaacttg agctttactt aaataacaaa gatgatcaaa
     2581 aaagatccat ctttcagaaa tcagagcgag gggggtttag gctgaaggag aatgtccagt
     2641 ttcatctgta catcagcacc tctccctgtg gagatgccag aatcttctca ccacatgagc
     2701 caatcctgga agaaccagca gatagacacc caaatcgtaa agcaagagga cagctacgga
     2761 ccaaaataga gtctggtgag gggacgattc cagtgcgctc caatgcgagc atccaaacgt
     2821 gggacggggt gctgcaaggg gagcggctgc tcaccatgtc ctgcagtgac aagattgcac
     2881 gctggaacgt ggtgggcatc cagggatccc tgctcagcat tttcgtggag cccatttact
     2941 tctcgagcat catcctgggc agcctttacc acggggacca cctttccagg gccatgtacc
     3001 agcggatctc caacatagag gacctgccac ctctctacac cctcaacaag cctttgctca
     3061 gtggcatcag caatgcagaa gcacggcagc cagggaaggc ccccaacttc agtgtcaact
     3121 ggacggtagg cgactccgct attgaggtca tcaacgccac gactgggaag gatgagctgg
     3181 gccgcgcgtc ccgcctgtgt aagcacgcgt tgtactgtcg ctggatgcgt gtgcacggca
     3241 aggttccctc ccacttacta cgctccaaga ttaccaaacc caacgtgtac catgagtcca
     3301 agctggcggc aaaggagtac caggccgcca aggcgcgtct gttcacagcc ttcatcaagg
     3361 cggggctggg ggcctgggtg gagaagccca ccgagcagga ccagttctca ctcacgccct
     3421 gcccaacttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     3481 agggcccgcg gttcgaaacc agcggaggcg gcggaagcgg gggaggaggc agcggcggag
     3541 gcggcagcgg gggcggcgga agcgctaccg gtaagcctat ccctaaccct ctcctcggtc
     3601 tcgattctac gcgtaccggt atggcttcta actttactca gttcgttctc gtcgacaatg
     3661 gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacggggtc gctgaatgga
     3721 tcagctctaa ctcgcgatca caggcttaca aagtaacctg tagcgttcgt cagagctctg
     3781 cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa agtggcaacc cagactgttg
     3841 gtggtgaaga gcttcctgta gccggatgga gatcttactt aaatatggaa ctaaccattc
     3901 caattttcgc cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa
     3961 aagatggaaa cccgattccc tcggccatcg cggccaactc cggcatctac ggaggtggag
     4021 gtggatctcg tacgggttag taatgagttt aaacccgctg atcagcctcg actgtgcctt
     4081 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4141 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4201 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4261 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4321 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4561 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     4621 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     4681 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     4741 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     4801 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     4861 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     4921 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     4981 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     5041 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     5101 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     5161 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     5221 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     5281 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     5341 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     5401 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     5461 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     5521 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     5581 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     5641 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     5701 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5761 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5881 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6001 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6781 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7081 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7201 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7261 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7321 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7561 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7621 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7801 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7981 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8041 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8101 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     8161 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     8221 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     8281 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     8341 tacgcgttga cattgattat tgactaggct tttgcaaaaa gctttgcaaa gatggataaa
     8401 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     8461 aatt
//
