LOCUS       sequence-214782-        7402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZSCAN18 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7402)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7402)
  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: pZSCAN18-donor
FEATURES             Location/Qualifiers
     source          1..7402
                     /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"
     regulatory      3634..3643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3884..3904
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3917..3961
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4001..4024
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4052..4054,4055..4057,4058..4771)
                     /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(4097..4118)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4358..4377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4809..5382,5383..5385,5386..5394)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4975..4992)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5202..5221
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5476..6279
                     /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(5539..5558)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6149..6168
                     /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 tcggtacccc cagcacacca acaagaacac ctagaggcgg
     2701 acttgggctg gagggccctg ggtgcctgtc gccagggggc ttgtcgctgg ggggtttggc
     2761 cttagctatt gaccccactc tcttcgctgg cagccgggcc cctccctcca ggagggtgcg
     2821 ctctgagcag gctctccttg catcgtgcat cttgtggcca agctagtggc caggtggaga
     2881 cctccaaagg cgcggtttcc agaacccggg ctcagtgggg ctgtccttcc tgtcctgtcg
     2941 gtggcctccg ttgttggagg cctccaggtc ccacatctgc agcagcaggc gcggggcatg
     3001 cgctgcaggg ctgcctggga gggacagcgc tgcttccaaa cagccgtgag ccccgcgggg
     3061 atccagagcc cggagccctg tgcgctgttc cggcagctga ggctcggctg ggtcctcctc
     3121 tccctggccc tgggcggggc gctcacgcta gggctctgac cttgttccag aaggcgggag
     3181 gcggcaggag agcgctgggt gcgcctgcga ggaggccgcc cccgcggggg tgctgcctga
     3241 gctgcctacg gaggcgcccc ctggggacgc ccttgccgat cccccgtcgg gcaccactga
     3301 ggaggaggaa gagcagcctg ggaaggcccc ggacccgcag gacccccagg acgcggagtc
     3361 cgactctgcc accggatcgc agaggcagtc cgtcatccag cagcctgccc cggacagggg
     3421 cacggcgaaa ctgggaacca agaggccgca ccccgaggat ggggacgggc agagcctcga
     3481 gggcgtctct agctccggcg acagcgcagg gctggaggcc gggcagggcc ctggggctga
     3541 cgagccgggc ttgtcccgcg ggaagcccta tgcctgcggc gagtgcgggg aggccttcgc
     3601 gtggctctcg cacctgatgg agcaccacag cagccatggc ggccggaagc gctacgcctg
     3661 tcagggctgc tggaagacct tccacttcag cctggcccta gccgagcacc agaagaccca
     3721 cgagaaggag aaaagctacg cgctgggggg cgcccggggc ccccaaccgt ccacccgcga
     3781 agcccaggcg ggggctaggg cgggcggtcc cccagagagc gtggagggcg aggctccccc
     3841 cgcaccccca gaggcgcaga gggggattcc aactactgca agcgagaatc tttattttca
     3901 gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc acatggacag
     3961 cggaggtgga ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc tgttccaggg
     4021 gcccctgtcg tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg agctgttcac
     4081 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4141 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4201 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     4261 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4321 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4381 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4441 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4501 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4561 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4621 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4681 agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg ccgccgggat
     4741 cactctcggc atggacgagc tgtacaagta atgatctaga gtcgagttaa ttaagaattc
     4801 cgcccccccc cctctccctc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4861 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4921 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4981 ccaaaggaat gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5041 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5101 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5161 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5221 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5281 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5341 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aacctgttac
     5401 attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta cataaacagt
     5461 aatacaaggg gtgttatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
     5521 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5581 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5641 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5701 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5761 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5821 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5881 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5941 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6001 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6061 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6121 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6181 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6241 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag tcgccgctgt gctgtcccgt
     6301 tccggagggg ccgctttgcc ggccgtgaat cccagacgag gcattgggcc tttccacgcc
     6361 cctgggtggc ggcttcctgt ggtgtttgtg gacgtcctct gcctgtgccc tgaatccgct
     6421 cctgaggcta agcgctccca acgagaaggg tccacgggaa gccctcacct ctgtaaacac
     6481 accctgggcc agcgctcgca tccgagggga gccgccggat gtggaagaag actcggcttt
     6541 cctgcagcca tttagtgccg ccccatgcta ggttatttga cattgtgcag tgtagagttg
     6601 ccttaaagtg cgtgatctgc cagtgctttc ttcaagtcac ccttgccccg attcctcctg
     6661 tttgcgctcc ccagggttgc tcaagtggaa attttgtcag ctgtttagcc ttttcgtact
     6721 tggcgtgatg tcaacttcac ttctaatctg caaaagcaga agctgtttcc tagtttacct
     6781 cgcgtgtgtt tacctatatg gagtagctcg cagagatcac agaaatgctt gcagcctaag
     6841 gcagggtttt cagaccgtgg gtcccagccc atttagtaaa atgggaaatc aattagcaag
     6901 tggtcaccag cattacacag caatgaagca gaataaagta ggccagaatg catcatgtag
     6961 taaaggcaaa tactgttttg tgaaactttt cacccataca tctaaatgtg agaactggtt
     7021 gcaatgtaag acatttcttg ctgggaagtt gtgagcaaaa taagttgaaa acactaataa
     7081 agatctgtct gtctgagcaa aggagactaa actccttggg ctacataagg tgatcttttc
     7141 attgattgca tttattctca agattacagt atcattccca ttgcctgcgc ataaacttaa
     7201 ctgctccctg cgcataaact taactgctcc ctgtagatac agaaactggt gcctcttttt
     7261 ctgtcactgt gtttgagcag gcagtctgtt gtaatgaccc ccaaatcaca gagacttagt
     7321 aagtacttat tcatgggccg ggcgtggtgg ctcacacctg taatcccaac actttgtgag
     7381 gctgaggcag gcagatcatg ga
//
