LOCUS       sequence-197865-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The wild-type Pum, fused to eGFP, and behind the ubiquitin 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Adamala KP, Martin-Alarcon DA, Boyden ES
  TITLE     Programmable RNA-binding protein composed of repeats of a single 
            modular unit.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2579-88. doi: 
            10.1073/pnas.1519368113. Epub 2016 Apr 26.
  PUBMED    27118836
REFERENCE   2  (bases 1 to 6754)
  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: 
            "10.1073/pnas.1519368113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-10- 10"; volume: "113"; issue: "19"; pages: 
            "E2579-88"
COMMENT     Sequence Label: [UBC][NRE Pum][eGFP]
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        711..1921
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1759..1778
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     promoter        1967..1985
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3143..3145,3146..3148,3149..3862)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3188..3209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3449..3468)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3796..3817
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3903..4024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3914..3933)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3968..3987
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4220..4675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4307..4326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4517..4538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4742..4761)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4861..4883
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4921..4939)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5007..5111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5112..5180,5181..5972)
                     /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(5330..5349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6143..6731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6632..6651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatggct cgacacgtct ccggatacgg
       61 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
      121 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
      181 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
      241 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
      301 tgtgtctgag cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt ggaagagctt
      361 gcctggagag atacagctgg gtcagtagga ctgggacagg cagctggaga attgccatgt
      421 agatgttcat acaatcgtca aatcatgaag gctggaaaag ccctccaaga tccccaagac
      481 caaccccaac ccacccaccg tgcccactgg ccatgtccct cagtgccaca tccccacagt
      541 tcttcatcac ctccagggac ggtgaccccc ccacctccgt gggcagctgt gccactgcag
      601 caccgctctt tggagaaggt aaatcttgct aaatccagcc cgaccctccc ctggcacaac
      661 gtaaggccat tatctctcat ccaactccag gacggagtca gtgaggatct ggcctccgcg
      721 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
      781 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
      841 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
      901 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
      961 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     1021 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     1081 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     1141 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagtccgc caagggctgt
     1201 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     1261 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     1321 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg
     1381 ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt
     1441 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
     1501 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
     1561 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
     1621 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
     1681 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
     1741 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
     1801 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
     1861 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
     1921 atgtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1981 ataggctagc ctcgagagcg cgcccgccac catgcgaggc atgggccgca gccgcctttt
     2041 ggaagatttt cgaaacaacc ggtaccccaa tttacaactg cgggagattg ccggacatat
     2101 aatggaattt tcccaagacc agcatgggtc cagattcatt cagctgaaac tggagcgtgc
     2161 cacaccagct gagcgccagc ttgtcttcaa tgaaatcctc caggctgcct accaactcat
     2221 ggtggatgtg tttggtaatt acgtcattca gaagttcttt gaatttggca gtcttgaaca
     2281 gaagctggct ttggcagaac ggattcgagg tcacgtcctg tcattggcac tacagatgta
     2341 tggctgccgt gttatccaga aagctcttga gtttattcct tcagaccagc agaatgagat
     2401 ggttcgggaa ctagatggcc atgtcttgaa gtgtgtgaaa gatcagaatg gcaattacgt
     2461 ggttcagaaa tgcattgaat gtgtacagcc ccagtctttg caatttatca tcgatgcgtt
     2521 taagggacag gtatttgcct tatccacaca tccttatggc tgccgagtga ttcagagaat
     2581 cctggagcac tgtctccctg accagacact ccctatttta gaggagcttc accagcacac
     2641 agagcagctt gtacaggatc aatatggaaa ttatgtaatc caacatgtac tggagcacgg
     2701 tcgtcctgag gataaaagca aaattgtagc agaaatccga ggcaatgtac ttgtattgag
     2761 tcagcacaaa tttgcaagca atgttgtgga gaagtgtgtt actcacgcct cacgtacgga
     2821 gcgcgctgtg ctcatcgatg aggtgtgcac catgaacgac ggtccccaca gtgccttata
     2881 caccatgatg aaggaccagt atgccaacta cgtggtccag aagatgattg acgtggcgga
     2941 gccaggccag cggaagatcg tcatgcataa gatccgaccc cacatcgcaa ctcttcgtaa
     3001 gtacacctat ggcaagcaca ttctggccaa gctggagaag tactacatga agaacggtgt
     3061 tgacttaggc ggacgcgggg gaggctcggg cggtggaagt ggaggcggtt caggcggtgg
     3121 gagtggaggc ggatcgggta ccatggtgag caagggcgag gagctgttca ccggggtggt
     3181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3241 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3301 gctgcccgtg ccctggccca ccctcgtgac caccttgacc tacggcgtgc agtgcttcgc
     3361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3421 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3541 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca aggtctatat
     3601 caccgccgac aagcagaaga acggcatcaa ggtgaacttc aagacccgcc acaacatcga
     3661 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3721 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3781 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3841 catggacgag ctgtacaagt gagtcgacgg ccggcccacg cggccgcttc gagcagacat
     3901 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3961 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4021 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     4081 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gatcggagga gacggagtgt
     4141 ggatccgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4201 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4261 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4321 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4381 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4441 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4501 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4561 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4621 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     4681 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     4741 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4801 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4861 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4921 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4981 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5041 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5101 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5161 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5221 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5281 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5341 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5401 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5461 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5521 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5581 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5641 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5701 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5761 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5821 agcgtgggtc gcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5881 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5941 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6001 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6061 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6361 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6421 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6481 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6541 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6601 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6661 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6721 gcctatggaa aaacgccagc aacgcggcct tttt
//
