LOCUS       sequence-376890-        8122 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 8122)
  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 8122)
  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: IGF2BP1
FEATURES             Location/Qualifiers
     source          1..8122
                     /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(3079..3103)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3228..3269
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3724..3741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3730..3954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4087..4106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4238..4285
                     /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             4293..5315
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4293..5315
                     /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    5445..5566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5482..5501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5536..5555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5615..5631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5615..5631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5628..5650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5639..5655
                     /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(5663..5669,5670..5687,5688..5693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5708..5729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5846..5863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6017..6605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6097..6116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6776..7567,7568..7636))
                     /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     7399..7418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7637..7741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7816..7835)
                     /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 gcagaagggc
     1321 cgtcaaggcc caccatgaac aagctttaca tcggcaacct caacgagagc gtgacccccg
     1381 cggacttgga gaaagtgttt gcggagcaca agatctccta cagcggccag ttcttggtca
     1441 aatccggcta cgccttcgtg gactgcccgg acgagcactg ggcgatgaag gccatcgaaa
     1501 ctttctccgg gaaagtagaa ttacaaggaa aacgcttaga gattgaacat tcggtgccca
     1561 aaaaacaaag gagccggaaa attcaaatcc gaaatattcc accccagctc cgatgggaag
     1621 tactggacag cctgctggct cagtatggta cagtagagaa ctgtgagcaa gtgaacaccg
     1681 agagtgagac ggcagtggtg aatgtcacct attccaaccg ggagcagacc aggcaagcca
     1741 tcatgaagct gaatggccac cagttggaga accatgccct gaaggtctcc tacatccccg
     1801 atgagcagat agcacaggga cctgagaatg ggcgccgagg gggctttggc tctcggggtc
     1861 agccccgcca gggctcacct gtggcagcgg gggccccagc caagcagcag caagtggaca
     1921 tcccccttcg gctcctggtg cccacccagt atgtgggtgc cattattggc aaggaggggg
     1981 ccaccatccg caacatcaca aaacagaccc agtccaagat agacgtgcat aggaaggaga
     2041 acgcaggtgc agctgaaaaa gccatcagtg tgcactccac ccctgagggc tgctcctccg
     2101 cttgtaagat gatcttggag attatgcata aagaggctaa ggacaccaaa acggctgacg
     2161 aggttcccct gaagatcctg gcccataata actttgtagg gcgtctcatt ggcaaggaag
     2221 gacggaacct gaagaaggta gagcaagata ccgagacaaa aatcaccatc tcctcgttgc
     2281 aagaccttac cctttacaac cctgagagga ccatcactgt gaagggggcc atcgagaatt
     2341 gttgcagggc cgagcaggaa ataatgaaga aagttcggga ggcctatgag aatgatgtgg
     2401 ctgccatgag cctgcagtct cacctgatcc ctggcctgaa cctggctgct gtaggtcttt
     2461 tcccagcttc atccagcgca gtcccgccgc ctcccagcag cgttactggg gctgctccct
     2521 atagctcctt tatgcaggct cccgagcagg agatggtgca ggtgtttatc cccgcccagg
     2581 cagtgggcgc catcatcggc aagaaggggc agcacatcaa acagctctcc cggtttgcca
     2641 gcgcctccat caagattgca ccacccgaaa cacctgactc caaagttcgt atggttatca
     2701 tcactggacc gccagaggcc caattcaagg ctcagggaag aatctatggc aaactcaagg
     2761 aggagaactt ctttggtccc aaggaggaag tgaagctgga gacccacata cgtgtgccag
     2821 catcagcagc tggccgggtc attggcaaag gtggaaaaac ggtgaacgag ttgcagaatt
     2881 tgacggcagc tgaggtggta gtaccaagag accagacccc tgatgagaac gaccaggtca
     2941 tcgtgaaaat catcggacat ttctatgcca gtcagatggc tcaacggaag atccgagaca
     3001 tcctggccca ggttaagcag cagcatcaga agggacagag taaccaggcc caggcacgga
     3061 ggaagtcagg cctcatgggc ccagctttct tgtacaaagt ggttgatatc cagcacagtg
     3121 gcggccgctc gagtctagag ggcccgcggt tcgaaaccag cggaggcggc ggaagcgggg
     3181 gaggaggcag cggcggaggc ggcagcgggg gcggcggaag cgctaccggt aagcctatcc
     3241 ctaaccctct cctcggtctc gattctacgc gtaccggtat ggcttctaac tttactcagt
     3301 tcgttctcgt cgacaatggc ggaactggcg acgtgactgt cgccccaagc aacttcgcta
     3361 acggggtcgc tgaatggatc agctctaact cgcgatcaca ggcttacaaa gtaacctgta
     3421 gcgttcgtca gagctctgcg cagaatcgca aatacaccat caaagtcgag gtgcctaaag
     3481 tggcaaccca gactgttggt ggtgaagagc ttcctgtagc cggatggaga tcttacttaa
     3541 atatggaact aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg
     3601 caatgcaagg tctcctaaaa gatggaaacc cgattccctc ggccatcgcg gccaactccg
     3661 gcatctacgg aggtggaggt ggatctcgta cgggttagta atgagtttaa acccgctgat
     3721 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3961 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4021 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4081 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4141 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4201 ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc cgaagttcct
     4261 attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg
     4321 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     4381 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     4441 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     4501 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     4561 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     4621 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     4681 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     4741 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     4801 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     4861 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     4921 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     4981 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5041 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     5101 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5161 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5221 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5281 gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat ttcgattcca
     5341 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5401 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5461 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5521 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5581 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5641 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5701 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5821 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7741 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7801 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7861 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     7921 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     7981 cgatgtacgg gccagatata cgcgttgaca ttgattattg actaggcttt tgcaaaaagc
     8041 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
     8101 ggtcttgaaa ggagtgggaa tt
//
