LOCUS       sequence-214787-        7034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor OVOL1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7034)
  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: pOVOL1-donor
FEATURES             Location/Qualifiers
     source          1..7034
                     /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             3564..3584
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3597..3641
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3681..3704
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3732..3734,3735..3737,3738..4451)
                     /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(3777..3798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4038..4057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4489..5062,5063..5065,5066..5074)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4655..4672)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4882..4901
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5156..5959
                     /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(5219..5238)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5829..5848
                     /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 tagaaggaca 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 tcggtaccca cgtcttccca atctgccagc ctgtggcccc
     2701 aacttctgga tgtctgtcag caatgccgtc ctcacccaca ggtgaccctt ggggacagtc
     2761 ccagtggaga cctgttcacc tgccgtgtct gccagaaggc cttcacctac cagcgcatgc
     2821 tgaaccgcca catgaagtgt cacaacgacg tcaagaggca cctctgcacg tactgcggga
     2881 agggcttcaa tgacaccttc gacctcaaga gacacgtccg aactcacact ggtaagtgga
     2941 agcccacggc tgggaggagc acgccggatc cgcctggccg cggccgtgcg ggaaaagtcc
     3001 agcaccccct gctctccctc ggtgacgggc agcctgggag ggcatcccgg agctcagggc
     3061 caaagtcctc ctgagctcat cagttaggag ttttgatttg agagtcactc gggaagttgg
     3121 tgttggacag gcctccgtcc atcccttggc cctagaggca ggggtcctgt cctttctgtg
     3181 tctggaagca gccgacagcc tctgaagtcc ctgccaggtc tctgatgctg gcccctgtcc
     3241 tccccacagg cgtgcggccc tacaagtgca gcctgtgtga caaggccttc acgcagcgct
     3301 gctctctgga gtctcacctc aagaagatcc atggtgtgca gcagaagtac gcgtacaagg
     3361 agcggcgggc caagctgtac gtgtgtgagg agtgcggctg cacatctgag agccaggagg
     3421 gccacgtcct gcacctgaag gagcaccacc ctgacagccc gctgctgcgc aagacctcca
     3481 agaaggtggc cgtggcacta cagaacactg tcacttccct gctgcagggc agcccccacc
     3541 tggggattcc aactactgca agcgagaatc tttattttca gggcgccgcc aaattcaaag
     3601 aaaccgctgc tgctaaattc gaacgccagc acatggacag cggaggtgga ggttcgagcg
     3661 gtccctcggg ttcgtcgagc ctggaagttc tgttccaggg gcccctgtcg tcgagcggtc
     3721 cctcgggttc gatggtgagc aaaggcgagg agctgttcac cggggtggtg cccatcctgg
     3781 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     3841 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3901 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3961 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4021 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4081 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4141 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4201 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4261 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4321 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     4381 atcacatggt ccttaaggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4441 tgtacaagta atgatctaga gtcgagttaa ttaagaattc cgcccccccc cctctccctc
     4501 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     4561 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     4621 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcagggtctg
     4681 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     4741 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     4801 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     4861 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4921 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     4981 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     5041 ctttgaaaaa cacgatgata atatggccac aacctgttac attgcacaag ataaaaatat
     5101 atcatcacga acagtaaaac tgtctgctta cataaacagt aatacaaggg gtgttatggg
     5161 atcggccatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5221 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5281 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5341 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5401 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5461 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5521 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5581 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5641 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5701 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5761 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5821 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5881 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5941 tcttgacgag ttcttctgag tcgtggctcg agccctgggg gtgctcctgg aagccccaag
     6001 agcatccagg attgcctccc agctgcctgg ccagcccacc ctcctgcaac ctctcacccg
     6061 aacaccagtg atcaggactg gagcccccgt gccttggtct cccccctggg cacacgtgct
     6121 cactcaggcc cagcaatgac ctctgctcat ttttgcattt ttgacttatg ggccgaggct
     6181 gttctgagcc tgggaagatg tacctatgtc aagagaaggg atgaggccaa ggctgccttc
     6241 aattagaagc agccgcccac agagacaggc actgtgtgcc tggcagcagg acttcctacc
     6301 cagaggaggt tcgagctagg atcccactgc ccccgcctct cagcacaggg caggggctgc
     6361 aggtccccag tggacatcag agtcaaaagc actggcaaag ggtacccctg caaacaactg
     6421 tggtgggggc tggcagcaga ccccccacct ggcagggctt ctaatgctca gggttctgga
     6481 gggctctgtc cttccggcaa ggagaggcac acatgtgtgc ccagccgtgt gtgtgcgtgt
     6541 gcttgtgtgt gtgcactgct gtgtgtgtgt gcacgcacag gaagcctttc cacatatcac
     6601 ctcatttcta agaaataaac tacaaggtgc caagaaggtt ttatttcctt ttatttttta
     6661 aagatgacaa atgtacagat gttaatatat ttttggtgcc aatggcgatg tttttaagag
     6721 tgggatggag ctggcttttc tccattcccg tgcgcttcta tttatcctgg acatttcaaa
     6781 cctcctctgt gccttggctc tgggcggggg ctgccccaac ccacccccgt tctttgtacg
     6841 tgctgagaca gccactagaa gatcttcctc cagcggcgcc ctggacggct gctcctgcga
     6901 acagcccatg gcatcttctg ctcttccctc ccggctctgc cctgcacatc ctgttgagcc
     6961 aagccccagt gacccggaga gctggcctga tgctgagagt gtgtcctcct ggggctttag
     7021 ggggcaggaa ggtg
//
