LOCUS       sequence-229691-        7172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to tag human transcription factor OTX1 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7172)
  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: pOTX1-donor
FEATURES             Location/Qualifiers
     source          1..7172
                     /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             3632..3661
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3842..3862
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3875..3919
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3959..3982
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4010..4012,4013..4015,4016..4729)
                     /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(4055..4076)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4316..4335)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4767..5340,5341..5343,5344..5352)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4933..4950)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5160..5179
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5434..6237
                     /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(5497..5516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6107..6126
                     /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 tcggtaccca agcagccaac agcagcaatt tagagatggc
     2701 agaacttctg actgggcaga ctgaaatatt ccgccctagg gcagagttgc ggtaggggtg
     2761 gggaggtgcg catccctgct ctggagccag ctgaggaggg gctttgagca gaaggatggg
     2821 ggagggtgga ggagctggga accgaggtag ggggcagggt ctggccaggc cagagacagg
     2881 aaggggcgga ggcctcggtg agaaaggatt gcgatattct ggaccgggag ttgggtccgc
     2941 ggggcggtgg agcaacaagc tcccctagct ccctttgacc cactctcccc catccggccc
     3001 actgcaggtc tggttcaaga accgccgcgc caaatgccgc cagcagcagc agagcgggag
     3061 cggaaccaag agccgcccag ccaagaagaa gtcctctcca gtgcgggaga gctcgggctc
     3121 cgaaagcagt ggccaattca cgccgccagc tgtgtccagc tctgcctcgt cctctagctc
     3181 ggcgtccagc tcttccgcca acccagcggc tgcagcggct gcgggactag gtgggaaccc
     3241 ggtggcggcc gcgtcgtcgc tgagtacacc agctgcctca tctatctgga gcccggcctc
     3301 catctcgcca ggctcagcgc ccgcgtccgt gtcggtgccg gagccattgg ccgcgcctag
     3361 caacacctcg tgtatgcagc gctccgtagc tgcaggcgcc gccaccgcag cagcctctta
     3421 tcccatgtcc tacggccagg gcggcagcta cggccaaggc taccctacgc cctcctcttc
     3481 ctactttggc ggcgtggact gcagctcata cctagcgccc atgcactcac atcaccaccc
     3541 gcaccagctc agccccatgg caccctcctc catggcgggc caccatcatc accacccaca
     3601 tgcgcaccac ccgttgagcc agtcctcagg ccaccaccac caccatcacc accaccacca
     3661 ccaaggctac ggtggctctg ggcttgcctt caactctgcc gactgcttgg attacaagga
     3721 gcctggcgcc gctgctgctt cctccgcctg gaaactcaac ttcaactccc ccgactgtct
     3781 ggactataag gaccaagcct catggcggtt ccaggtcttg gggattccaa ctactgcaag
     3841 cgagaatctt tattttcagg gcgccgccaa attcaaagaa accgctgctg ctaaattcga
     3901 acgccagcac atggacagcg gaggtggagg ttcgagcggt ccctcgggtt cgtcgagcct
     3961 ggaagttctg ttccaggggc ccctgtcgtc gagcggtccc tcgggttcga tggtgagcaa
     4021 aggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     4081 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     4141 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4201 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4261 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4321 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4381 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4441 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4501 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4561 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4621 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc ttaaggagtt
     4681 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat gatctagagt
     4741 cgagttaatt aagaattccg cccccccccc tctccctccc cccccctaac gttactggcc
     4801 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     4861 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     4921 ggggtctttc ccctctcgcc aaaggaatgc agggtctgtt gaatgtcgtg aaggaagcag
     4981 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     5041 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     5101 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     5161 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     5221 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     5281 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     5341 atggccacaa cctgttacat tgcacaagat aaaaatatat catcacgaac agtaaaactg
     5401 tctgcttaca taaacagtaa tacaaggggt gttatgggat cggccattga acaagatgga
     5461 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5521 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5581 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5641 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5701 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5761 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5821 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5881 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5941 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6001 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6061 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6121 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6181 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagtc
     6241 tgagcccagg aatgaaagag gagaagaaac gcaactacct gcgccctccg tggtcccgat
     6301 cctgttgctg ctgctgcacc gcccgccttt gcctcgtctt ctccaaaact gaattttcac
     6361 cccccaaaaa gatgtccatt gcctgcactg ccgcccccat ttttgtgcca cttgcttggg
     6421 gggatgtgca aacccaccct gccccttgga tggggggacc ggtgcttcgg cttggcctac
     6481 acattctata caggagagat gtattatttc cccccttcag cccctactaa actcttaagc
     6541 ctccccttcc agtctttctg gacagctatt aagcacttgc agccttcgga ggctctgcgc
     6601 tctgatccgc tgtttgagcc caacacttta atttattctt tctggacact ggagtcacta
     6661 actggcgtgt ttctgcccat tggagcaccc gcacactact ccaaatcaaa accactaaga
     6721 gttcctcccg cgcagactgc tgccccttca gctgccctcg attttgctcc acgcctgccg
     6781 gccagagcct cccggcgttt cttccgcccc agcggagtgc gctggggcgc gccagggcta
     6841 ggcccgccgg aggagcgcgt ccccagcctt ccgcgcacag agccgcatcc cgccccgccc
     6901 tgcgctggac tggttcaagc ttccgcctcg gcgggaacgc tgtacatagt caggtccgtt
     6961 ccagggacca cttaaacttt ttagttgctg ttggttggtt gaactgaaca tatcttgtct
     7021 tagcacccag gaaacagaac tttaagatat atacagcaca tatatatata tatacatata
     7081 tatatacaca tatataaaaa acaaaagcaa aaaatatttt ccctctgtcc gtcccccttc
     7141 tgctcttcct caatcaatgg cgcttttttt tt
//
