LOCUS       sequence-211704-        8033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor JUNB with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8033)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 8033)
  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: pJUNB-donor
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..89)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(71..89)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(96..113)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(96..112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(105..127)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             250..552
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     431..450
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             901..1695
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             901..1695
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      1896..1905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1902..2276
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      2414..3002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2903..2922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3156..3173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3290..3311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3326..3331,3332..3349,3350..3356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3364..3380
                     /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)."
     primer_bind     3369..3391
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3388..3404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3388..3404
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3422..3440
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3422..3439
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4572..4592
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4605..4649
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4689..4712
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4740..4742,4743..4745,4746..5459)
                     /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(4785..4806)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5046..5065)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(5497..6070,6071..6073,6074..6082)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5663..5680)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5890..5909
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6164..6967
                     /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"
ORIGIN
        1 aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
       61 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
      121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagccta
      241 tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
      301 gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt
      361 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
      421 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
      481 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
      541 tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta gatccttttc
      601 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
      661 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
      721 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
      781 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt ctcgccgcca
      841 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
      901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     1021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     1081 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     1141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     1201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     1261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     1321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     1381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     1441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     1501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     1681 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     1741 catctgtgcg gtatttcaca ccgcatacag gtggcacttt tcggggaaat gtgcgcggaa
     1801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1861 cctgataaat gcttcaataa tagcacgtga ggagggccac catggccaag ttgaccagtg
     1921 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     1981 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     2041 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     2101 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     2161 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     2221 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     2281 tgctaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2521 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3001 aaaacgccag caacgcggcc tttttacggt tcctgggctt ttgctggcct tttgctcaca
     3061 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3181 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3241 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3301 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3361 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     3421 tatttaggtg acactataga atactcaagc tatgcatcaa gcttggtacc gagctcggat
     3481 ccactagtaa cggccgccag tgtgctggaa ttcgccctta acagcccttc taccacgacg
     3541 actcatacac agctacggga tacggccggg cccctggtgg cctctctcta cacgactaca
     3601 aactcctgaa accgagcctg gcggtcaacc tggccgaccc ctaccggagt ctcaaagcgc
     3661 ctggggctcg cggacccggc ccagagggcg gcggtggcgg cagctacttt tctggtcagg
     3721 gctcggacac cggcgcgtct ctcaagctcg cctcttcgga gctggaacgc ctgattgtcc
     3781 ccaacagcaa cggcgtgatc acgacgacgc ctacaccccc gggacagtac ttttaccccc
     3841 gcgggggtgg cagcggtgga ggtgcagggg gcgcaggggg cggcgtcacc gaggagcagg
     3901 agggcttcgc cgacggcttt gtcaaagccc tggacgatct gcacaagatg aaccacgtga
     3961 caccccccaa cgtgtccctg ggcgctaccg gggggccccc ggctgggccc gggggcgtct
     4021 acgccggccc ggagccacct cccgtttaca ccaacctcag cagctactcc ccagcctctg
     4081 cgtcctcggg aggcgccggg gctgccgtcg ggaccgggag ctcgtacccg acgaccacca
     4141 tcagctacct cccacacgcg ccgcccttcg ccggtggcca cccggcgcag ctgggcttgg
     4201 gccgcggcgc ctccaccttc aaggaggaac cgcagaccgt gccggaggcg cgcagccggg
     4261 acgccacgcc gccggtgtcc cccatcaaca tggaagacca agagcgcatc aaagtggagc
     4321 gcaagcggct gcggaaccgg ctggcggcca ccaagtgccg gaagcggaag ctggagcgca
     4381 tcgcgcgcct ggaggacaag gtgaagacgc tcaaggccga gaacgcgggg ctgtcgagta
     4441 ccgccggcct cctccgggag caggtggccc agctcaaaca gaaggtcatg acccacgtca
     4501 gcaacggctg tcagctgctg cttggggtca agggacacgc cttcgattat gatattccaa
     4561 ctactgcaag cgagaatctt tattttcagg gcgccgccaa attcaaagaa accgctgctg
     4621 ctaaattcga acgccagcac atggacagcg gaggtggagg ttcgagcggt ccctcgggtt
     4681 cgtcgagcct ggaagttctg ttccaggggc ccctgtcgtc gagcggtccc tcgggttcga
     4741 tggtgagcaa aggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4801 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4861 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4921 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4981 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     5041 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     5101 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     5161 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     5221 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     5281 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     5341 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     5401 ttaaggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     5461 gatctagagt cgagttaatt aagaattccg cccccccccc tctccctccc cccccctaac
     5521 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     5581 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     5641 agcattccta ggggtctttc ccctctcgcc aaaggaatgc agggtctgtt gaatgtcgtg
     5701 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     5761 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     5821 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     5881 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     5941 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     6001 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     6061 cgatgataat atggccacaa cctgttacat tgcacaagat aaaaatatat catcacgaac
     6121 agtaaaactg tctgcttaca taaacagtaa tacaaggggt gttatgggat cggccattga
     6181 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6241 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6301 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     6361 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6421 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6481 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6541 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6601 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6661 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6721 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6781 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6841 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6901 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6961 cttctgaacg tcccctgccc ctttacggac accccctcgc ttggacggct gggcacacgc
     7021 ctcccactgg ggtccaggga gcaggcggtg ggcacccacc ctgggaccta ggggcgccgc
     7081 aaaccacact ggactccggc cctcctaccc tgcgcccagt ccttccacct cgacgtttac
     7141 aagccccccc ttccactttt ttttgtatgt tttttttctg ctggaaacag actcgattca
     7201 tattgaatat aatatatttg tgtatttaac agggagggga agagggggcg atcgcggcgg
     7261 agctggcccc gccgcctggt actcaagccc gcggggacat tgggaagggg acccccgccc
     7321 cctgccctcc cctctctgca ccgtactgtg gaaaagaaac acgcacttag tctctaaaga
     7381 gtttatttta agacgtgttt gtgtttgtgt gtgtttgttc tttttattga atctatttaa
     7441 gtaaaaaaaa aattggttct ttattaattt ctgttgtctt tttttccaag ctgggagggc
     7501 ggggggaaaa aaaaaagcac tggtttgccc ccagctcagt gctgttggtg gctcggtcct
     7561 gtatgtgtcc ccctcgtcgg ttcggcgcag gcatcttgtg gtcccagccc aggagtccca
     7621 cccttcccgc gtccccagat ctccagggtt ggatggttgg ggcgcgggac gcggtccagg
     7681 gacccaggag ctgaaggcag ggtgctccgg ccgagacttg gagtgcgcag gcgcgtcccc
     7741 gcccagccgc gcgcgccggg gctttccccg ctgacgcagc ggaagcgctg cccatacaag
     7801 gaccgattct gcccagtgac gcgaccgcgg tctctgggca gattccggga atcccctccc
     7861 ccgctctgcc gggcagggcc gcggcgccgg gaagggggcc gggattttcc cgggcaggcg
     7921 gctgcccggg cacggaaagc cccaggcctg ggtccagagc gcccgcggtg ggcgggcagc
     7981 gctcctgggc tccgggagcc acccctgtgc caccttcaag acactggcgc cta
//
