LOCUS       sequence-229833-        7204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor ZNF397 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7204)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7204)
  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: pZNF397-donor
FEATURES             Location/Qualifiers
     source          1..7204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..49)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(33..49)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(46..68)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    57..73
                     /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(81..87,88..105,106..111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    126..147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(264..281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(435..1023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(515..534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1194..1985,1986..2054))
                     /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     1817..1836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2055..2159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2227..2245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2283..2305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2405..2424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2618..2640
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2632..2649
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2633..2649
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3880..3900
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3913..3957
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3997..4020
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4048..4050,4051..4053,4054..4767)
                     /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(4093..4114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4354..4373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4805..5378,5379..5381,5382..5390)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4971..4988)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5198..5217
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5472..6275
                     /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(5535..5554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6145..6164
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gacctgcagg catgcagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
       61 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
      121 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
      181 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      241 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      301 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      361 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      601 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      841 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1201 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1261 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1321 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1381 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1441 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1501 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1561 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1621 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1681 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1741 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1801 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1861 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1921 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1981 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2041 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2101 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2161 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2221 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     2281 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2341 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     2401 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     2461 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     2521 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2581 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2641 acggccagtg aattcgagct cggtacccac cctcttttgg ccagttgaaa aaaaagatac
     2701 tggaggacag tgacatcttt gtaggtagaa gcttctgttt tctctcatgt agaaagtttt
     2761 gacggtgaca agagaaatgt aatatctgtt tttttgctac ttattgtttc agattgtgag
     2821 aacagtgaaa cagcaacaaa agagggcatc tcagaagaaa aatcacaggg actccctcag
     2881 gaaccttcat ttcgaggaat tagtgagcat gaaagcaatt tagtgtggaa gcaaggaagt
     2941 gctacagggg agaaactaag atctccttcc caagggggca gttttagtca agtgatcttc
     3001 acaaacaaat ctctaggaaa gagagacctt tatgatgagg ctgaaagatg cttgattcta
     3061 actacagact ctataatgtg tcagaaagtt cctccagaag agagacctta tagatgtgat
     3121 gtatgtgggc acagcttcaa gcagcattcc tctctaacac aacatcagag aatccatact
     3181 ggagaaaagc cctataaatg taaccagtgt gggaaggcct ttagtttgag gtcctatctt
     3241 attattcatc agagaattca tagtggtgag aaagcatatg aatgtagtga atgtgggaaa
     3301 gctttcaatc agagctcagc cctcattaga catcggaaaa tccatactgg tgagaaagct
     3361 tgtaaatgta atgagtgtgg caaagcattc agtcaaagtt catatctcat tatacatcaa
     3421 agaattcaca ctggtgagaa accttatgag tgtaatgaat gtgggaaaac ctttagccag
     3481 agctcaaaac tcattagaca tcagcgaatt cacacaggag agagacccta tgaatgtaat
     3541 gaatgtggaa aagctttcag gcagagctca gagctgatta ctcatcagag aatacatagt
     3601 ggagagaaac cctatgaatg tagtgaatgt ggaaaagctt tcagtttgag ctcaaacctt
     3661 atcagacatc agaggattca tagtggggag gaaccttatc agtgtaatga atgtggcaaa
     3721 actttcaaaa ggagctcagc ccttgttcag catcagagaa ttcattctgg ggatgaagct
     3781 tatatatgta atgaatgtgg gaaggctttc aggcacagat cggtccttat gcgccatcaa
     3841 agagtccaca ctataaaggg gattccaact actgcaagcg agaatcttta ttttcagggc
     3901 gccgccaaat tcaaagaaac cgctgctgct aaattcgaac gccagcacat ggacagcgga
     3961 ggtggaggtt cgagcggtcc ctcgggttcg tcgagcctgg aagttctgtt ccaggggccc
     4021 ctgtcgtcga gcggtccctc gggttcgatg gtgagcaaag gcgaggagct gttcaccggg
     4081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4141 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     4261 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4321 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     4441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     4501 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     4561 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4621 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     4681 cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc cgggatcact
     4741 ctcggcatgg acgagctgta caagtaatga tctagagtcg agttaattaa gaattccgcc
     4801 cccccccctc tccctccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     4861 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4921 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     4981 aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5041 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5101 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5161 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     5221 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     5281 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     5341 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc tgttacattg
     5401 cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata aacagtaata
     5461 caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt tctccggccg
     5521 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5581 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5641 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5701 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5761 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5821 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5881 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5941 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6001 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6061 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6121 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6181 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6241 tcgccttcta tcgccttctt gacgagttct tctgagtctt tgtgaatact gtgaatagtg
     6301 taaatacttc agtcagattt ttaagtttgt tagtcaaaag agtttacttt ggagcaaaac
     6361 tccataaagg ttataaaata ctaggtcttg agtctagctt gctttgtgca gcatttccca
     6421 gtgctaatgt aaagtgtccc ttgaaagctt ttcctgtgac taatcagaac agaatacaga
     6481 agaatcatta cttccagctc ttctcttatt aggaatactc aggaaatacg aaaagtggga
     6541 atgtaacatt gaaacctcat tttgtatgaa agtgtcatga atatagcaac ccaggctctg
     6601 tcactgcatc tgattgttag tgtgccaggt tatggggctt gtgtggactg tgtgaggcac
     6661 ttcgactcag gaactaaaaa aaaaaaaaaa ctgacccaat aaagaacagt gacagagcag
     6721 caggaaaggt ggggaaatgg cttcatcaat gatttgttga gcccagggca ggccaggaat
     6781 tagataggca tgagaagaaa gaatataatt tacccaaagg ttattttttg agtacctact
     6841 ttgtcccagg aactaggaat acagtggaaa acaagactgt agtctctaca gtctaatggg
     6901 caaggcagcc agacaggcag tgaaagtgga atgctcctat gtgaccttaa ggagcacctg
     6961 actcagcctt ggataaggaa atgggggaaa tggcctgaag gatgaggagg agttagccaa
     7021 ggtggtactc tagggaagtg gtaccccagt aaagaacagt agccaaaggc cagaaacaaa
     7081 gtgtggagca ttccttgagt tcctgttgta gtgaggtctt gtctgtcaga gaagtctggg
     7141 tctggccaga ctcttaagca gtgccaatgg agaagttcct gttaatagac aaaacctgga
     7201 gccc
//
