LOCUS       sequence-241248-        7401 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 7401)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7401)
  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..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1000..1009
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1250..1270
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1283..1327
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1367..1390
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(1418..1420,1421..1423,1424..2137)
                     /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(1463..1484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1724..1743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(2175..2748,2749..2751,2752..2760)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2341..2358)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2568..2587
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2842..3645
                     /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
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLFDEFF"
     primer_bind     complement(2905..2924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3515..3534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4802..4818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4802..4818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4815..4837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4826..4842
                     /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(4850..4856,4857..4874,4875..4880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4895..4916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5033..5050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5204..5792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5284..5303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5963..6754,6755..6823))
                     /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     6586..6605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6824..6928)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6996..7014
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7052..7074)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7174..7193
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7387..7401,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7401,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgagctcg gtacccccag cacaccaaca agaacaccta
       61 gaggcggact tgggctggag ggccctgggt gcctgtcgcc agggggcttg tcgctggggg
      121 gtttggcctt agctattgac cccactctct tcgctggcag ccgggcccct cctccaggag
      181 ggtgcgctct gagcaggctc tccttgcatc gtgcatcttg tggccaagct agtggccagg
      241 tggagacctc caaaggcgcg gtttccagaa cccgggctca gtggggctgt ccttcctgtc
      301 ctgtcggtgg cctccgttgt tggaggcctc caggtcccac atctgcagca gcaggcgcgg
      361 ggcatgcgcc gcagggctgc ctgggaggga cagcgctgct tccaaacagc cgtgagcccc
      421 gcggggatcc agagcccgga gccctgtgcg ctgttccggc agctgaggct cggctgggtc
      481 ctcctctccc tggccctggg cggggcgctc acgctagggc tctgaccttg ttccagaagg
      541 cgggaggcgg caggagagcg ctgggtgcgc ctgcgaggag gccgcccccg cgggggtgct
      601 gcctgagctg cctacggagg cgccccctgg ggacgccctt gccgatcccc cgtcgggcac
      661 cactgaggag gaggaagagc agcctgggaa ggccccggac ccgcaggacc cccaggacgc
      721 ggagtccgac tctgccaccg gatcgcagag gcagtccgtc atccagcagc ctgccccgga
      781 caggggcacg gcgaaactgg gaaccaagag gccgcacccc gaggatgggg acgggcagag
      841 cctcgagggc gtctctagct ccggcgacag cgcagggctg gaggccgggc agggccctgg
      901 ggctgacgag ccgggcttgt cccgcgggaa gccctatgcc tgcggcgagt gcggggaggc
      961 cttcgcgtgg ctctcgcacc cgatggagca ccacagcagc catggcggcc ggaagcgcta
     1021 cgcctgtcag ggctgctgga agaccttcca cttcagcctg gccctagccg agcaccagaa
     1081 gacccacgag aaggagaaaa gctacgcgct ggggggcgcc cggggccccc aaccgtccac
     1141 ccgcgaagcc caggcggggg ctagggcggg cggtccccca gagagcgtgg agggcgaggc
     1201 tccccccgca cccccagagg cgcagagggg gattccaact actgcaagcg agaatcttta
     1261 ttttcagggc gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat
     1321 ggacagcgga ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt
     1381 ccaggggccc ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct
     1441 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1501 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1561 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1621 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1681 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1741 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1801 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1861 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1921 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1981 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     2041 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     2101 cgggatcact ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa
     2161 gaattccgcc cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg
     2221 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2281 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2341 ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2401 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2461 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2521 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2581 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2641 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2701 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     2761 tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata
     2821 aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt
     2881 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2941 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3001 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3061 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3121 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3181 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3241 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3301 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3361 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3421 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3481 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3541 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3601 tcgcagcgca tcgccttcta tcgccttttt gacgagttct tctgagtcgc cgctgtgctg
     3661 tcccgttccg gaggggccgc tttgccggcc gtgaatccca gacgaggcat tgggcctttc
     3721 cacgcccctg ggtggcggct tcctgtggtg tttgtggacg tcctctgcct gtgccctgaa
     3781 tccgctcctg aggctaagcg ctcccaacga gaagggtcca cgggaagccc tcacctctgt
     3841 aaacacaccc tgggccagcg ctcgcatccg aggggagccg ccggatgtgg aagaagactc
     3901 ggctttcctg cagccattta gtgccgcccc atgctaggtt atttgacatt gtgcagtgta
     3961 gagttgcctt aaagtgcgtg atctgccagt gctttcttca agtcaccctt gccccgattc
     4021 ctcctgtttg cgctccccag ggttgctcaa gtggaaattt tgtcagctgt ttagcctttt
     4081 cgtacttggc gtgatgtcaa cttcactcct aatctgcaaa aacagaagct gtttcctagt
     4141 ttacctcgcg tgtgtttacc tatatggagt agctcgcaga gatcacagaa atgcttgcag
     4201 cctaaggcag ggttttcaga ccgtgggtcc cagcccattt agtaaaatgg gaaatcaatt
     4261 agcaagcggt caccagcatt acacagcaat gaagcagaat aaagtaggcc agaatgcatc
     4321 atgtagtaaa ggcaaatact gttttgtgaa acttttcacc catacatcta aatgtgagaa
     4381 ctggttgcaa tgtaagacat ttcttgctgg gaagttgtga gcaaaataag ttgaaaacac
     4441 taataaagat ctgtctgtct gagcaaagga gactaaactc cttgggctac ataaggtgat
     4501 cttttcattg attgcattta ttctcaagat tacagtatca ttcccattgc ctgcgcataa
     4561 acttaactgc tccctgcgca taaacttaac tgctccctgt agatacagaa actggtgcct
     4621 ctttttctgt cactgtgttt gagcaggcag tctgttgtaa tgacccccaa atcacagaga
     4681 cttagtaagt acttattcat gggccgggcg tggtggctca cacctgtaat cccaacactt
     4741 tgtgaggctg aggcaggcag atcatggaga cctgcaggca tgcaagcttg gcgtaatcat
     4801 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4861 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5641 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6961 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     7021 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     7081 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7141 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     7201 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     7261 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     7321 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     7381 ggttttccca gtcacgacgt t
//
