LOCUS       sequence-471720-        8810 bp    DNA     circular SYN 06-MAY-2025
DEFINITION  For overexpression of mEGFP-NUP98-DDX10-KS-FtoG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8810)
  TITLE     Hnisz Lab plasmids 2025
REFERENCE   2  (bases 1 to 8810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRK5_mEGFP-NUP98-DDX10-KS-FtoG
FEATURES             Location/Qualifiers
     source          1..8810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      934..943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(940..942,943..945,946..1656)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(985..1006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1246..1265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1593..1614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(4754..4780)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     regulatory      5110..5119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5123..5257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5214..5233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5326..5683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5326..5517
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5349..5369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5534..5669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5703..5720)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5703..5719)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5712..5734)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5932..6387
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6019..6038)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6229..6250
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6404..6423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6523..6545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6583..6601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6669..6773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6774..6842,6843..7634)
                     /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(6992..7011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7805..8393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8294..8313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8547..8564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8681..8702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8717..8722,8723..8740,8741..8747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8755..8771
                     /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)."
     primer_bind     8760..8782
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8779..8795
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8779..8795
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtaccg gtcgccacca tggtgagcaa gggcgaggag
      961 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1021 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1081 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1141 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1201 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1261 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1321 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1381 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1441 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1501 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1561 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1621 gccgggatca ctctcggcat ggacgagctg tacaagaccg gcggcatgtt taacaaatca
     1681 tttggaacac cctttggggg tggcacaggt ggctttggca caacttcaac atttggacag
     1741 aatactggct ttggcactac tagtggaggg gcatttggaa catctgcatt tggttctagc
     1801 aacaatactg gaggcctctt tggaaattca cagactaaac caggaggatt gtttggaacc
     1861 agttcattta gccagccagc tacctccaca agcactggct ttgggtttgg tacgtcaaca
     1921 ggaacagcaa ataccttgtt tggaactgca agcacaggga ccagtctctt ctcatcccaa
     1981 aacaatgcct ttgcacaaaa taaaccaact ggctttggca attttggaac cagtactagc
     2041 agtggaggac tctttggaac cacaaatacc acctctaatc cttttggcag cacatctggc
     2101 tccctctttg ggccaagtag ttttacagct gctcctactg ggactactat taaatttaac
     2161 cctccaactg gtacagatac tatggtcaaa gctggagtta gcactaacat aagtaccaag
     2221 caccagtgta ttactgctat gaaagaatat gaaagcaagt cactagagga acttcgttta
     2281 gaggattatc aggctaacag gaagggccca cagaaccagg tgggagcagg taccacaact
     2341 ggcttgtttg ggtcttctcc agccacttcc agcgcaacag gactcttcag ctcctccacc
     2401 actaattcag gctttgcata tggtcagaac aaaactgcct ttggaactag tacaactgga
     2461 tttggaacaa atccaggtgg tctctttggc caacagaatc agcagactac cagcctcttc
     2521 agcaaaccat ttggccaggc tacaaccacc cagaacactg gcttttcctt tggtaatacc
     2581 agcaccatag gacagccaag caccaacacc atgggattat ttggagtaac ccaagcctca
     2641 cagcctggag gtctttttgg gacagctaca aacaccagca ctgggacagc atttggaaca
     2701 ggaacaggtc tctttgggca gaccaatact ggatttggtg ctgttggttc gaccctgttt
     2761 ggcaataaca agcttactac atttggaagc agcacaacca gtgcaccttc atttggtaca
     2821 accagtggcg ggctctttgg ttttggcaca aataccagtg ggaatagtat ttttggaagt
     2881 aaaccagcac ctgggactct tggaactggg cttggtgcag gatttggaac agctcttggt
     2941 gctggacagg catctttgtt tgggaacaac caacctaaga ttggagggcc tcttggtaca
     3001 ggagcctttg gggcccctgg atttaatact acgacagcca ctttgggctt tggagccccc
     3061 caggccccag tagttcttga tgaagcagat agaatcttgg atatgggctt tgctgatacc
     3121 atgaatgctg ttattgaaaa tctccccaag aaacgtcaga ctttactttt ctcagcaaca
     3181 caaactaaat ctgtaaagga ccttgcacgc ttgagtttga aaaaccctga gtatgtctgg
     3241 gttcatgaaa aagcaaaata tagcacccct gccactttgg aacagaacta catagtctgt
     3301 gagctgcagc aaaaaataag tgtgctgtat tcctttttga gaagccatct gaagaagaag
     3361 agcattgtat ttttttccag ttgcaaagag gtccagtatc tgtaccgagt gttttgccgg
     3421 ctacgtcctg gtgtttctat ccttgcactc catggtcgac agcagcaaat gagaagaatg
     3481 gaagtctata atgagtttgt ccgtaagaga gctgcagtac tctttgctac tgatattgca
     3541 gccaggggtc tggatttccc ggccgtgaat tgggttcttc agtttgattg tcctgaggat
     3601 gccaacacat atattcacag agcaggtaga actgccaggt acaaagagga tggtgaagct
     3661 ttgctaattt tgctaccctc agaaaaagct atggtgcagc agcttcttca gaagaaagta
     3721 cctgtgaagg aaatcaaaat caatccagaa aaacttatag atgtccagaa aaaattggaa
     3781 tctattttag ctcaagatca agatttaaaa gaaagagctc aaaggtgttt cgtctcctat
     3841 gtacgatctg tatatctgat gaaggataaa gaagtatttg atgtgagcaa gttacctata
     3901 cctgaatatg ccctgtctct tggtcttgct gtggcaccac gcgtaagatt tcttcagaaa
     3961 atgcagaaac aacccaccaa agaattggta aggagccaag ccgataaagt aattgagcca
     4021 agggctccct ccctcaccaa tgacgaagtg gaagaattta gagcctactt caatgagaaa
     4081 atgtccatcc ttcaaaaagg tggaaaaaga ctcgaaggga cagagcacag acaggataat
     4141 gatactggta atgaagaaca ggaagaagaa gaagacgatg aagaagaaat ggaagagaaa
     4201 ctggcaaaag caaaaggatc tcaagcccca tctcttccta acaccagtga ggcacagaag
     4261 atcaaggaag ttcctacaca gttcttggac agagatgagg aggaagaaga tgctgatttc
     4321 ttgaaggtga agcggcataa tgtgtttgga ttggacctta aagacgagaa aacattacag
     4381 aagaaagaac cttctaaatc cagcatcaag aaaaaaatga ccaaagttgc agaagcaaaa
     4441 aaagtaatga agagaaattt taaagtgaat aagaagataa catttactga tgaaggggag
     4501 ttggttcagc agtggccaca aatgcagaaa tctgccatca aggatgctga ggaagatgat
     4561 gacacaggtg gtatcaactt acataaagca aaggaaagac ttcaggaaga ggacaaattt
     4621 gacaaagaag aatataggaa aaaaattaag gcaaagcatc gggagaaaag actgaaagaa
     4681 agggaagcca gaagagaagc caacaagaga caagcaaagg ccaaagatga agaggaagcc
     4741 tttctggatt ggagtgatga tgatgatgat gatgatgatg gatttgatcc aagcacactc
     4801 ccagatccag ataaatacag aagctctgaa gattcagata gtgaagatat ggaaaataaa
     4861 ataagtgata ccaagaagaa gcaggggatg aagaagagga gcaacagtga agtggaagac
     4921 gtgggaccaa caagtcataa cagaaagaag gccaggtggg acactttaga gcctttggat
     4981 accggcctgt ctttagcaga ggatgaagag ctggtgttac atctgctaag aagtcaaagc
     5041 atgatgatga tgaataagtt ggttctagcg cagggtggtg gttcctgtct atgaacctgc
     5101 agaagcttgg ccgccatggc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattaattcg gcgcagcacc
     5281 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     5341 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctgt taacagcttg
     5701 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5761 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5821 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     5881 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     5941 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6001 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6061 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6121 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     6181 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6241 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     6301 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6361 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     6421 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6481 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6541 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     6601 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6661 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6721 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6781 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6841 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6901 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6961 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7021 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7081 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7141 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7201 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7261 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7321 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7381 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7441 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7501 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7561 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7621 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7681 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7741 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7801 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7861 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7921 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7981 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8041 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8101 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8161 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8221 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8281 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8341 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8401 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8461 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8521 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8581 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     8641 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     8701 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8761 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattaa
//
