LOCUS       sequence-228532-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor FOXA3 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7169)
  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: pFOXA3-donor
FEATURES             Location/Qualifiers
     source          1..7169
                     /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             complement(3671..3682)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3887..3907
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3920..3964
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4004..4027
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4055..4057,4058..4060,4061..4774)
                     /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(4100..4121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4361..4380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4812..5385,5386..5388,5389..5397)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4978..4995)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5205..5224
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5479..6282
                     /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(5542..5561)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6152..6171
                     /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 tcggtacccg agcatggtgg gtgggaaagc agggggcaga
     2701 aagggcgtgt ccagggagtg aggagagacc ttgagagact gctttctaca gatagaggta
     2761 atgatggaag ggatttgtag agaaggcaag atggacagac agacggacac tcagtgggtc
     2821 ctgggatcct ttgctgacca gcagagtgga gccccactga cccctccttt catctttccc
     2881 ctaggtctac tcgccggtga ccccagtgcc caccatggcc cccctcaact cctacatgac
     2941 cctgaatcct ctaagctctc cctatccccc tggggggctc cctgcctccc cactgccctc
     3001 aggacccctg gcacccccag cacctgcagc ccccctgggg cccactttcc caggcctggg
     3061 tgtcagcggt ggcagcagca gctccgggta cggggccccg ggtcctgggc tggtgcacgg
     3121 gaaggagatg ccgaaggggt atcggcggcc cctggcacac gccaagccac cgtattccta
     3181 tatctcactc atcaccatgg ccatccagca ggcgccgggc aagatgctga ccttgagtga
     3241 aatctaccag tggatcatgg acctcttccc ttactaccgg gagaatcagc agcgctggca
     3301 gaactccatt cgccactcgc tgtctttcaa cgactgcttc gtcaaggtgg cgcgttcccc
     3361 agacaagcct ggcaagggct cctactgggc cctacacccc agctcaggga acatgtttga
     3421 gaatggctgc tacctgcgcc gccagaaacg cttcaagctg gaggagaagg tgaaaaaagg
     3481 gggcagcggg gctgccacca ccaccaggaa cgggacaggg tctgctgcct cgaccaccac
     3541 ccccgcggcc acagtcacct ccccgcccca gcccccgcct ccagcccctg agcctgaggc
     3601 ccagggcggg gaagatgtgg gggctctgga ctgtggctca cccgcttcct ccacacccta
     3661 tttcactggc ctggagctcc caggggagct gaagctggac gcgccctaca acttcaacca
     3721 ccctttctcc atcaacaacc taatgtcaga acagacacca gcacctccca aactggacgt
     3781 ggggtttggg ggctacgggg ctgaaggtgg ggagcctgga gtctactacc agggcctcta
     3841 ttcccgctct ttgcttaatg catccgggat tccaactact gcaagcgaga atctttattt
     3901 tcagggcgcc gccaaattca aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga
     3961 cagcggaggt ggaggttcga gcggtccctc gggttcgtcg agcctggaag ttctgttcca
     4021 ggggcccctg tcgtcgagcg gtccctcggg ttcgatggtg agcaaaggcg aggagctgtt
     4081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4681 caaagacccc aacgagaagc gcgatcacat ggtccttaag gagttcgtga ccgccgccgg
     4741 gatcactctc ggcatggacg agctgtacaa gtaatgatct agagtcgagt taattaagaa
     4801 ttccgccccc ccccctctcc ctcccccccc ctaacgttac tggccgaagc cgcttggaat
     4861 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4921 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4981 tcgccaaagg aatgcagggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5041 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5101 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5161 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5221 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5281 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5341 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaacctgt
     5401 tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc ttacataaac
     5461 agtaatacaa ggggtgttat gggatcggcc attgaacaag atggattgca cgcaggttct
     5521 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5581 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5641 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5701 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5761 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5821 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5881 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5941 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6001 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6061 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6121 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6181 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6241 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagtccaggg gttgggaaca
     6301 tggtggtggg tatggctgga gctcacacca cgaagctctt ggggcctgat ccttctggtg
     6361 acacttcact tgtcccattg gttaacatct gggtgggtct attacttact gtgatgactg
     6421 ctgtctcagt gggcatggtg ttgatccacg gggtactgtg ataaccacca tggatacatt
     6481 ttggtggccc actgggtact gtgaggactg ctacattgat ggatgttatt ggctaatcca
     6541 ctgcatggtt tgatggccac catctcggtt ggccctttgg gtgtgatggt gatagcattt
     6601 cagtgacatc ttctttggcc ccccccatta ggtgctgtgc ccacttcttt tttggtgtac
     6661 ttggcacagt aggtgccaag ttggccacca ttctgtgtaa cacctttttt ggcccattgg
     6721 gtgctttgat ggacatcata ctgggtaggt gacaacgtca gtgggccacc atgtgccatg
     6781 atggctgctg cagccccgtg ttggccatgt cgtcaccatt ctctctggca tgggttgggt
     6841 aggggatgga ggtgagaata ctccttggtt ttctctgaag cccacccttt cccccaactc
     6901 tggtccagga gaaaccagaa aaggctggtt agggtgtggg gaatttctac tgaagtctga
     6961 ttctttcccg ggaagcgggg tactggctgt gtttaatcat taaaggtacc gtgtccgcct
     7021 cttactgaat gtctcttttc cttggccagt gggccctgtg gggcttgaac acctgaattt
     7081 ctgctcctct gtcccttctg cttcctgccc ctaagagctg gctgcctctg ggggaggagt
     7141 tgggggaggc atcgacctga cctttggct
//
