LOCUS       sequence-230195-        7328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor TFAP2B with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7328)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7328)
  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: pTFAP2B-donor
FEATURES             Location/Qualifiers
     source          1..7328
                     /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             3877..3897
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3910..3954
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3994..4017
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4045..4047,4048..4050,4051..4764)
                     /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(4090..4111)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4351..4370)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4802..5375,5376..5378,5379..5387)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4968..4985)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5195..5214
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5469..6272
                     /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(5532..5551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6142..6161
                     /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 tcggtaccct gccagccatt tgaccttcag ggctagaacc
     2701 atcccatcca gctcctatat taaaatttct caaataagct caatccatag gtaggcacgc
     2761 aggatttcta aggctgggag attcctctaa attgtaggtg gcttgaggaa ataaaaacct
     2821 tcaacaaata aaaatctatc ctccctagtg tcttaacagc ctacctttag gttttattta
     2881 aactatatgg gaatcctttt atattgacaa cagtctggtt atacaaagat tttcagtgcc
     2941 ctacacttgg aagaaagaaa ggagggaaag gcagaaacta gctccctccc ccaatactgc
     3001 aggtaaacag gtttttgctt tgaatggaat cagcattctc tagaataaac agtttcttgc
     3061 cctgggccat ggaaggctcc agcaatctgc ttcagtgcca gggacaggct tgtggccgag
     3121 gggcctaaat ctttcagggc tttttgcgcc tgatcaccaa gaggttggag aggtgggagg
     3181 ggcgcggggg aaacagctca aaagaaatgc caggtaagag ctcaatatta tgaagtatta
     3241 aaaatcaacc gctaaagcct agcgaatgtc ccaaagcgag tccaggcggt cctggaatgg
     3301 aggggagaaa ggctgaagtc gggggaaggc aaggctttgg ggtggaaggg gtgcattacc
     3361 tccccacccc cgctttcctt tcagtcccag ctgctcgaga agggagggcg ctgggaaagg
     3421 aaacagagcg gaatcgccca cattagcctc gctcttcggt gacccggcgc ctctgggctt
     3481 gtgtgagcgt ctcctttcta atgccaatga caacgacact gagggtgatt ttctgtgcct
     3541 cgcccacccc tttgcaggca actttgtaaa gaatttacgg atctactggc gcaggaccgg
     3601 acaccgatag ggaacagccg acccagcccc atcctggagc cggggatcca gagctgcctc
     3661 acgcacttca gcctcatcac gcacggcttc ggcgccccgg ccatttgcgc cgcgctcacg
     3721 gccctgcaga actatctcac cgaggcgctc aaaggcatgg acaagatgtt cttgaacaac
     3781 accaccacta acaggcacac gtctggggaa ggcccaggta gtaaaactgg cgacaaggag
     3841 gagaaacaca ggaaagggat tccaactact gcaagcgaga atctttattt tcagggcgcc
     3901 gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcggaggt
     3961 ggaggttcga gcggtccctc gggttcgtcg agcctggaag ttctgttcca ggggcccctg
     4021 tcgtcgagcg gtccctcggg ttcgatggtg agcaaaggcg aggagctgtt caccggggtg
     4081 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4141 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4201 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     4261 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4321 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4381 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4441 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     4501 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     4561 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4621 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4681 aacgagaagc gcgatcacat ggtccttaag gagttcgtga ccgccgccgg gatcactctc
     4741 ggcatggacg agctgtacaa gtaatgatct agagtcgagt taattaagaa ttccgccccc
     4801 ccccctctcc ctcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     4861 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     4921 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     4981 aatgcagggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     5041 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     5101 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     5161 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5221 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     5281 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     5341 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacctgt tacattgcac
     5401 aagataaaaa tatatcatca cgaacagtaa aactgtctgc ttacataaac agtaatacaa
     5461 ggggtgttat gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt
     5521 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5581 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5641 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5701 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5761 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5821 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5881 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5941 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6001 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6061 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6121 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6181 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6241 ccttctatcg ccttcttgac gagttcttct gagtctgaaa aatttttaaa aaaagaagga
     6301 aaaatgtttt aaatacaaaa ggaaaaacag acaaaaattt aattttagct ttaaaatatt
     6361 ggattggctt tggaagaatt atattaggta gaatacacat acaatcaaaa tttaaaaaaa
     6421 aaaagctaaa taacttaaaa aaaaactgag gcgtacaacg gagcaacaat atcggttctc
     6481 agtgtctatt tcaagataca tttggagaca accgtccgga ttttccactt cggttctttc
     6541 gagtttagta atactgataa taaaagaaaa ccatgatttc cccttccctt tggaaaataa
     6601 acataagact aaacatgaga aaaacgctaa cttattggaa gaaaatcgga gaaacgttgg
     6661 tgtcaatgct ttgagagctg gttgactgag acgcacgaac tttttaattt taaatatatt
     6721 tttaggaaac tctcgcagtc cccgccctcc atctcacctc acccgtctcc caaccaccct
     6781 tttccatgtt accctccctt cctcacattg ttacgggaat cttctgggat gaaaatgagt
     6841 gtggttggcc cttttgcgtt gtttcagtcc tctcggtacc tccccgccca gccctgcgat
     6901 cttaactcac cgggcccggc tccccggccg cttgcataat tagggagggc gagggcgggg
     6961 cgggaggcct gtggagacca ggcggaggct gcaatttttg gtgccggccg gcagagcagg
     7021 ttctggcggc tgaggaaaat ccggaccaat aagttgattc aaacatcatc agttcctttc
     7081 agaaatgtta ctagctccca gccttgccag catctgcata gagagaatct cacatttatt
     7141 atatttgtgt catctactaa ttttatgaac tatagtaaaa aaaaaaaaca cacacacaca
     7201 atatgaatac gttgattagt atgtaattcc ttcggagggg tatgtaccat ttcattctct
     7261 tctgttttcc aaagctttgc ccgcatggtt tatgagcttc ttcaaagagg acttgggctt
     7321 cctacaca
//
