LOCUS       sequence-178839-        6752 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 6752)
  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 6752)
  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..6752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        719..1928
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1765..1784
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     promoter        1973..1991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3149..3151,3152..3154,3155..3868)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3194..3215)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3455..3474)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3802..3823
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3909..4030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3920..3939)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3974..3993
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4226..4681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4523..4544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4748..4767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4867..4889
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4927..4945)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5013..5117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5118..5186,5187..5978)
                     /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(5336..5355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6149..6737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6638..6657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acacgtctcc
       61 ggatacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct
      121 agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc
      181 gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct
      241 ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaagc
      301 tttaggcttg tgtctgagcc tgcatgtttg atggtgtctg gatgcaagca gaaggggtgg
      361 aagagcttgc ctggagagat acagctgggt cagtaggact gggacaggca gctggagaat
      421 tgccatgtag atgttcatac aatcgtcaaa tcatgaaggc tggaaaagcc ctccaagatc
      481 cccaagacca accccaaccc acccaccgtg cccactggcc atgtccctca gtgccacatc
      541 cccacagttc ttcatcacct ccagggacgg tgaccccccc acctccgtgg gcagctgtgc
      601 cactgcagca ccgctctttg gagaaggtaa atcttgctaa atccagcccg accctcccct
      661 ggcacaacgt aaggccatta tctctcatcc aactccagga cggagtcagt gaggatctgg
      721 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc
      781 acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg
      841 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
      901 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
      961 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     1021 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     1081 ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc
     1141 cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agtccgccaa
     1201 gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag
     1261 caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt
     1321 cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc
     1381 taatgcggga aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac
     1441 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     1501 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     1561 cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt
     1621 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     1681 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     1741 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     1801 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     1861 gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     1921 tgttagacgt gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga
     1981 ctcactatag gctagcctcg agggcgcgcc cgccaccatg cgaggcatgg gccgcagccg
     2041 ccttttggaa gattttcgaa acaaccggta ccccaattta caactgcggg agattgccgg
     2101 acatataatg gaattttccc aagaccagca tgggtccaga ttcattcagc tgaaactgga
     2161 gcgtgccaca ccagctgagc gccagcttgt cttcaatgaa atcctccagg ctgcctacca
     2221 actcatggtg gatgtgtttg gtaattacgt cattcagaag ttctttgaat ttggcagtct
     2281 tgaacagaag ctggctttgg cagaacggat tcgaggtcac gtcctgtcat tggcactaca
     2341 gatgtatggc tgccgtgtta tccagaaagc tcttgagttt attccttcag accagcagaa
     2401 tgagatggtt cgggaactag atggccatgt cttgaagtgt gtgaaagatc agaatggcaa
     2461 ttacgtggtt cagaaatgca ttgaatgtgt acagccccag tctttgcaat ttatcatcga
     2521 tgcgtttaag ggacaggtat ttgccttatc cacacatcct tatggctgcc gagtgattca
     2581 gagaatcctg gagcactgtc tccctgacca gacactccct attttagagg agcttcacca
     2641 gcacacagag cagcttgtac aggatcaata tggaaattat gtaatccaac atgtactgga
     2701 gcacggtcgt cctgaggata aaagcaaaat tgtagcagaa atccgaggca atgtacttgt
     2761 attgagtcag cacaaatttg caagcaatgt tgtggagaag tgtgttactc acgcctcacg
     2821 tacggagcgc gctgtgctca tcgatgaggt gtgcaccatg aacgacggtc cccacagtgc
     2881 cttatacacc atgatgaagg accagtatgc caactacgtg gtccagaaga tgattgacgt
     2941 ggcggagcca ggccagcgga agatcgtcat gcataagatc cgaccccaca tcgcaactct
     3001 tcgtaagtac acctatggca agcacattct ggccaagctg gagaagtact acatgaagaa
     3061 cggtgttgac ttaggcggac gcgggggagg ctcgggcggt ggaagtggag gcggttcagg
     3121 cggtgggagt ggaggcggat cgggtaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3181 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3241 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3301 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg
     3361 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3421 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3481 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3541 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
     3601 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
     3661 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3721 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     3781 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3841 tctcggcatg gacgagctgt acaagtgagt cgacggccgg cccacgcggc cgcttcgagc
     3901 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3961 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4021 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     4081 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgatc ggaggagacg
     4141 gagtgtggat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4201 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4441 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4561 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4621 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     4681 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4741 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4801 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4861 cagtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgccagac
     4921 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4981 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5221 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5281 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5341 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5401 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5461 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5521 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5641 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5701 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5761 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5821 ccggtgagcg tgggtcgcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5941 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6001 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6061 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6721 ggcggagcct atggaaaaac gccagcaacg cg
//
