LOCUS       sequence-229895-        7400 bp    DNA     circular SYN 15-APR-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor TSHZ2 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pTSHZ2-donor
FEATURES             Location/Qualifiers
     source          1..7400
                     /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             3882..3902
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3915..3959
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3999..4022
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4050..4052,4053..4055,4056..4769)
                     /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(4095..4116)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4356..4375)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4837..5389
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4981..4998)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5208..5227
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5482..6285
                     /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(5545..5564)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6155..6174
                     /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 aaacacctcc agaagccaaa aagaccgagc
     2701 tgggtcccct gaaggaggag gagaagctga tgaaagaggg cagcgagaag gagaaacccc
     2761 agcccctgga gcccacatct gctctgagca atgggtgcgc cctcgccaac cacgccccgg
     2821 ccctgccatg catcaaccca ctcagcgccc tgcagtccgt cctgaacaat cacttgggca
     2881 aagccacgga gcccttgcgc tcaccttcct gctccagccc aagttcaagc acaatttcca
     2941 tgttccacaa gtcgaatctc aatgtcatgg acaagccggt cttgagtcct gcctccacaa
     3001 ggtcagccag cgtgtccagg cgctacctgt ttgagaacag cgatcagccc attgacctga
     3061 ccaagtccaa aagcaagaaa gccgagtcct cgcaagcaca atcttgtatg tccccacctc
     3121 agaagcacgc tctgtctgac atcgccgaca tggtcaaagt cctccccaaa gccaccaccc
     3181 caaagccagc ctcctcctcc agggtccccc ccatgaagct ggaaatggat gtcaggcgct
     3241 ttgaggatgt ctccagtgaa gtctcaactt tgcataaaag aaaaggccgg cagtccaact
     3301 ggaatcctca gcatcttctg attctacaag cccagtttgc ctcgagcctc ttccagacat
     3361 cagagggcaa atacctgctg tctgatctgg gcccacaaga gcgtatgcaa atctctaagt
     3421 ttacgggact ctcaatgacc actatcagtc actggctggc caacgtcaag taccagctta
     3481 ggaaaacggg cgggacaaaa tttctgaaaa acatggacaa aggccacccc atcttttatt
     3541 gcagtgactg tgcctcccag ttcagaaccc cttctaccta catcagtcac ttagaatctc
     3601 acctgggttt ccaaatgaag gacatgaccc gcttgtcagt ggaccagcaa agcaaggtgg
     3661 agcaagagat ctcccgggta tcgtcggctc agaggtctcc agaaacaata gctgccgaag
     3721 aggacacaga ctctaaattc aagtgtaagt tgtgctgtcg gacatttgtg agcaaacatg
     3781 cggtaaaact ccacctaagc aaaacgcaca gcaagtcacc cgaacaccat tcacagtttg
     3841 taacagacgt ggatgaagaa gggattccaa ctactgcaag cgagaatctt tattttcagg
     3901 gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac atggacagcg
     3961 gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct ggaagttctg ttccaggggc
     4021 ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa aggcgaggag ctgttcaccg
     4081 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4141 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4201 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     4261 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4321 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4381 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4441 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4501 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4561 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4621 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4681 accccaacga gaagcgcgat cacatggtcc ttaaggagtt cgtgaccgcc gccgggatca
     4741 ctctcggcat ggacgagctg tacaagtaat gatctagagt cgagttaatt aagaattccg
     4801 cccccccccc tctccctccc cccctccccc cccctaacgt tactggccga agccgcttgg
     4861 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     4921 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     4981 ctctcgccaa aggaatgcag ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     5041 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     5101 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     5161 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     5221 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     5281 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     5341 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5401 tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc tgcttacata
     5461 aacagtaata caaggggtgt tatgggatcg gccattgaac aagatggatt gcacgcaggt
     5521 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5581 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5641 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5701 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5761 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5821 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5881 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5941 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6001 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6061 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6121 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6181 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6241 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcct ctgcaggtat
     6301 gggtttgctc tgaggcattg cgattagcct ggtgaggagc tttcttacag ggagatgggt
     6361 ctgcttagag gcagctagca tctcccaatg ccaagcagga tagtagcttg ctggagtagg
     6421 atttatttta atgaaagacc agaacatgaa aactaagtcc taatcattct cttaagtcct
     6481 ctagtttaga gctgggaact agaataaaac gggtttagag agataaagtt cacagattac
     6541 tactgctttt tttaggcctt catcatccat aatcctgaat tacccaaggc aagctccagt
     6601 gtccagccaa ggctatcttt cattttaatt attagatact aacttctgtc accattccat
     6661 ttcaccaccc cacctttcca tagcaatgcc aatgacttat aaagtgacaa aaagttgagc
     6721 aatggaagaa agattggaga gtgctagaag tagttactat atagagattc cttaatcagc
     6781 tcatgaatag cttaaagttg tctttatgat tgttacttgt aatgaatgct tttttgcagc
     6841 ttacttttag taattgccac cttccacctc ttccctttgg ttattttcta cattaagtgt
     6901 cctatgaatg agcatgtgcc cttaacaggt tggtagaaaa atattcttct ttgcagaaca
     6961 ttgagatgaa aagaagctta ttttcttctt tcatcttttg gttttcttgt ggatagacca
     7021 gtattagagc ccacctttaa caaagataga ggcaatgtgt gcccttgagg tttcatttca
     7081 cataaagaaa caaatggctt tctgtacact tatgttaatg tgcatggagt cccatagctt
     7141 gaagataatg tgcatcatgt gtcatagaag ttccatgcct tatatagtca accatgccca
     7201 gaccatttgt gtcattttta ggaaacaggg cagagaccaa ggtcaaatgc tttggcagat
     7261 tagtttcacc agccattttc aaatgttgag gataagcaag gctgggtctt aaccaatctc
     7321 tttcatttgc ccatttgtgt tttctctact tctttttctc tcccttccag catgatgccc
     7381 gcatacacta agggccctcc
//
