LOCUS       sequence-290396-        7029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsAsb9-3UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7029)
  AUTHORS   Bhandari D, Raisch T, Weichenrieder O, Jonas S, Izaurralde E
  TITLE     Structural basis for the Nanos-mediated recruitment of the CCR4-NOT 
            complex and translational repression.
  JOURNAL   Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113.
  PUBMED    24736845
REFERENCE   2  (bases 1 to 7029)
  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: 
            "10.1101/gad.237289"; journalName: "Genes Dev"; date: "2014-04-15- 
            15"; volume: "28"; issue: "8"; pages: "888-901"
COMMENT     Sequence Label: pCIneo-RLuc-HsAsb9-3UTR_X
FEATURES             Location/Qualifiers
     source          1..7029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1089..1098
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1095..2030
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2733..2854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2744..2763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2798..2817
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3040..3495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3127..3146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3337..3358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3612..3969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3612..3803
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3635..3655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3820..3955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4020..4814
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4074..4093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4684..4703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4878..4926
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4967..4986)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5086..5108
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5146..5164)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5232..5336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5337..5405,5406..6197)
                     /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     complement(5555..5574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6368..6956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6857..6876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc caccatggct tccaaggtgt acgaccccga gcaacgcaaa cgcatgatca
     1141 ctgggcctca gtggtgggct cgctgcaagc aaatgaacgt gctggactcc ttcatcaact
     1201 actatgattc cgagaagcac gccgagaacg ccgtgatttt tctgcatggt aacgctgcct
     1261 ccagctacct gtggaggcac gtcgtgcctc acatcgagcc cgtggctaga tgcatcatcc
     1321 ctgatctgat cggaatgggt aagtccggca agagcgggaa tggctcatat cgcctcctgg
     1381 atcactacaa gtacctcacc gcttggttcg agctgctgaa ccttccaaag aaaatcatct
     1441 ttgtgggcca cgactggggg gcttgtctgg cctttcacta ctcctacgag caccaagaca
     1501 agatcaaggc catcgtccat gctgagagtg tcgtggacgt gatcgagtcc tgggacgagt
     1561 ggcctgacat cgaggaggat atcgccctga tcaagagcga agagggcgag aaaatggtgc
     1621 ttgagaataa cttcttcgtc gagaccatgc tcccaagcaa gatcatgcgg aaactggagc
     1681 ctgaggagtt cgctgcctac ctggagccat tcaaggagaa gggcgaggtt agacggccta
     1741 ccctctcctg gcctcgcgag atccctctcg ttaagggagg caagcccgac gtcgtccaga
     1801 ttgtccgcaa ctacaacgcc taccttcggg ccagcgacga tctgcctaag atgttcatcg
     1861 agtccgaccc tgggttcttt tccaacgcta ttgtcgaggg agctaagaag ttccctaaca
     1921 ccgagttcgt gaaggtgaag ggcctccact tcagccagga ggacgctcca gatgaaatgg
     1981 gtaagtacat caagagcttc gtggagcgcg tgctgaagaa cgagcagtaa ttctagatct
     2041 gtgacatccc ttggggctct agtagaaggg cccccttctt tgatgcagtt atgccgcctt
     2101 agaattcgga agtgttttgg aatccagcag catcataaga taaccaaact cgtcctccca
     2161 gaggatctga aacagtttct cctacatctt taaatgcatc tagggaatgg attcacaaac
     2221 gatgtgaaaa cattattgag tgttgtagcc actagaattt taaaatcaag ttggatttat
     2281 agagtttgac tagttttttc gattagattt gtatttgtta taaacttgtt tatggagttt
     2341 gactaatttt ttctattcaa tttgtatttg ttaaactcaa gccagggttg aaagacactg
     2401 catacgtttg tattattagt tagaaggcat gaagactttt ttccctgcat ggagagtgtc
     2461 ataagttatt gttttgcata tctactgcat gccaagcact ttctgcatca tctaatttag
     2521 ccctcacagc cactgggtca agatgtccaa ttttccagag taaggataga ggagtcaaat
     2581 tcaaatacag gttttctgac attaacttat gtgatgactt gatcgaggca ggcttttcca
     2641 gtatcactgt cctggttcca tctctgctat atgggaagaa ctcgaggcgg ccgcttccct
     2701 ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2761 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2821 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2881 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2941 tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat
     3001 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3181 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3481 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3541 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3601 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3901 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3961 ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct agagccacca
     4021 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4081 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4141 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4201 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4261 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4321 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4381 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4441 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4501 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4561 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4621 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4681 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4741 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4801 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4861 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     4921 tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca atctgctctg
     4981 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5041 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5101 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5161 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5341 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5401 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5461 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5521 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5581 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5641 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5701 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5761 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5821 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5881 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5941 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6001 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6061 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6121 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6481 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
     7021 gacagatct
//
