LOCUS       sequence-229726-        7115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ELF4 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7115)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7115)
  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"
COMMENT     Sequence Label: pELF4-donor
FEATURES             Location/Qualifiers
     source          1..7115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..50)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(47..69)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    58..74
                     /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(82..88,89..106,107..112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(265..282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(436..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(516..535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2284..2306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2406..2425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2619..2641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2633..2650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2634..2650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3691..3711
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3724..3768
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3808..3831
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3859..3861,3862..3864,3865..4578)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3904..3925)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4165..4184)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4616..5189,5190..5192,5193..5201)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4782..4799)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5009..5028
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5283..6086
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5346..5365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5956..5975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
       61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2641 gacggccagt gaattcgagc tcggtacccg ctgcagtgag cagtaatcgt gtcactgcac
     2701 tccatcccgg gcagcagagt gagaccctgt ctctagaaaa agaaaaaaac aacaaacagc
     2761 aaataaatcg acaaacacct tcagctccat tcctctcgtg ccctccttcc taaccagctg
     2821 agggacactt cctgccttaa ccaactctgt tctctgtttc ttctcagtgc atcttcagtg
     2881 cccagcaaca tccacctagg agtggccccc gtggggtcgg gctcggccct gaccctgcag
     2941 acgatcccac tgaccacggt gctgaccaat gggcctcctg ccagtactac tgctcccact
     3001 cagctcgttc tccagagtgt tccagcggcc tctactttca aggacacctt cactttgcag
     3061 gcctctttcc ccctgaacgc cagtttccaa gacagccagg tggcagcccc aggggctcca
     3121 ctgattctca gtggcctccc ccaacttctg gctggggcca accgtccgac caacccggcg
     3181 ccacccacgg tcacaggggc tggaccagca gggcccagct ctcagccccc tgggactgtc
     3241 attgctgcct tcatcaggac ttctggcact acagcagccc ctagggtcaa ggaggggcca
     3301 ctgaggtcct cctcctatgt tcagggtatg gtgacggggg cccccatgga ggggctgctg
     3361 gttcctgaag agaccctgag ggagctcctg agagatcagg ctcatcttca gccacttcca
     3421 acccaggtgg tttccagggg ttcccacaat ccgagccttc tgggcaacca gactttgtct
     3481 cctcccagcc gccccactgt tgggctgacc ccagtggctg aacttgagct ctcctcaggc
     3541 tcagggtccc tgctgatggc tgagcctagt gtgaccacat ctgggagcct tctgacaaga
     3601 tcccccaccc cagccccttt ctccccattc aaccctactt ccctcattaa gatggagccc
     3661 catgacatag ggattccaac tactgcaagc gagaatcttt attttcaggg cgccgccaaa
     3721 ttcaaagaaa ccgctgctgc taaattcgaa cgccagcaca tggacagcgg aggtggaggt
     3781 tcgagcggtc cctcgggttc gtcgagcctg gaagttctgt tccaggggcc cctgtcgtcg
     3841 agcggtccct cgggttcgat ggtgagcaaa ggcgaggagc tgttcaccgg ggtggtgccc
     3901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4021 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     4081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4261 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4321 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4441 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     4501 aagcgcgatc acatggtcct taaggagttc gtgaccgccg ccgggatcac tctcggcatg
     4561 gacgagctgt acaagtaatg atctagagtc gagttaatta agaattccgc ccccccccct
     4621 ctccctcccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     4681 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4741 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4801 gggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4861 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4921 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4981 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     5041 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5101 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5161 ggttttcctt tgaaaaacac gatgataata tggccacaac ctgttacatt gcacaagata
     5221 aaaatatatc atcacgaaca gtaaaactgt ctgcttacat aaacagtaat acaaggggtg
     5281 ttatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5341 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5401 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5461 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5521 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5581 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5641 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5701 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5761 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5821 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5881 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5941 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6001 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6061 atcgccttct tgacgagttc ttctgagtct aagcaaaggg gtcagggcaa gtgtgaccca
     6121 ccaggcaaaa ttgagcagca ttttcatagg gaccgacttc agtagcacac ctgcccctgc
     6181 atttcagtgg gatgtcaata cacttgaccc caagtccccc ggccctgcct ggtgtcactg
     6241 tggccaaaca gtgcccagct taagcatccc tggcatcaga ctatggcctt caagagcact
     6301 agggcatatg cttttggcag cataacgggc tgacttggtg atggagggaa aaagccttga
     6361 gccaggcaga agtttgtggc cagggtttgt gcagcagctt tgtgagaaga gcccttctac
     6421 ctggctctat ctcactggct gcattcccta cacagggaat ttactaccct atatgtgaat
     6481 atccctgtat gtacttgtgt gtacttgttg gtctgtatct tagtttcttt ggggaggaca
     6541 gggctgtagc tgtgaggtct tgtctccaag ggtgtgtgta tgtctccgtg gatcagccac
     6601 agggataggg attttgtttt taagggaaag cattctctaa ttccctttgt tcatgccgag
     6661 attcagttgc tctgagacta tggggtacaa gtttgatcct ccgaatctgg agatgttgta
     6721 gagctggaac gagtgcagag taggaacgct ttgatgcgca tgcacattgg ggaagatgcg
     6781 ctcctcaggg acacaaaggc cgagtggggt aaaaccacga agggagggaa gggaagtcag
     6841 ctctgggagc agccctcact ggctggacca aggtactctt cctggagttt gccgtgttag
     6901 caaccacagt caccttgcag tcaggctgga atcttgggcc accccaaagt gctttgctga
     6961 aggatttaga cggggatgaa gtgccctcca gcctcagagc tagccacaaa gcccccagag
     7021 ctgaattcat tgagtatttg tgcctagggc ttgggctgtt tgtgtgatac cggccccccg
     7081 ccagacaata gcctttgctg acaccccagc ctact
//
