LOCUS       sequence-124818-        7469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF5-TF with mCherry marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7469)
  AUTHORS   Slomovic S, Collins JJ
  TITLE     DNA sense-and-respond protein modules for mammalian cells.
  JOURNAL   Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
  PUBMED    26389572
REFERENCE   2  (bases 1 to 7469)
  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"; journalName: "Nat 
            Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585."
COMMENT     Sequence Label: pVITRO1-SS-127
FEATURES             Location/Qualifiers
     source          1..7469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(74..94)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        256..1562
                     /label=mEF-1-alpha promoter
                     /note="strong constitutive promoter for mouse elongation 
                     factor EF-1-alpha"
     intron          583..1530
                     /label=mEF-1-alpha intron
                     /note="intron upstream of the start codon of mouse 
                     EF-1-alpha"
     CDS             1577..1600
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1601..1621
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1901..2050
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     polyA_signal    2082..2203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2093..2112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2147..2166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2391..2979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2880..2899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3059..3362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3477..4782
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3819..4750
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             4795..5505
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWQASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4940..4958)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5208..5227
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5512..5956
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5989..6035
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6027..6045
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6055..6849
                     /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             6055..6849
                     /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"
     primer_bind     complement(6109..6128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6719..6738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6896..7468
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
ORIGIN
        1 cctgcagggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
       61 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
      121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
      181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      241 ccactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc aaggggagag
      301 cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc cacgacagcg
      361 cgaggagcat gcgctcaggg ctgagcgcgg ggagagcaga gcacacaagc tcatagaccc
      421 tggtcgtggg ggggaggacc ggggagctgg cgcggggcaa actgggaaag cggtgtcgtg
      481 tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc ggcagtcgcc
      541 ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagggg cgggtgtggc
      601 ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg ccgggccccg ctgtcgtcgg
      661 cggggattag ctgcgagcat tcccgcttcg agttgcgggc ggcgcgggag gcagagtgcg
      721 aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc
      781 cgcgccgccg ccgcgtgcta ctccggccgc actctggtct tttttttttt tgttgttgtt
      841 gccctgctgc cttcgattgc cgttcagcaa taggggctaa caaagggagg gtgcggggct
      901 tgctcgcccg gagcccggag aggtcatggt tggggaggaa tggagggaca ggagtggcgg
      961 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgg
     1021 ggtttttccc gaagcaacca ggctggggtt agcgtgccga ggccatgtgg ccccagcacc
     1081 cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc cctaactagg gtgaggccat
     1141 cccgtccggc accagttgcg tgcgtggaaa gatggccgct cccgggccct gttgcaagga
     1201 gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg
     1261 aagagggcct ggtccctcac cggctgctgc ttcctgtgac cccgtggtcc tatcggccgc
     1321 aatagtcacc tcgggctttt gagcacggct agtcgcggcg gggggagggg atgtaatggc
     1381 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     1441 catttttgga atttgtcccc ttgagttttg agcggagcta attctcgggc ttcttagcgg
     1501 ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa aaccaccgct aattcaaagc
     1561 aaccggtgat atcatggact acaaggacga cgatgacaag cccaagaaaa agcgcaaggt
     1621 ctctagaccc ggggagcgcc ccttccagtg tcgcatttgc atgcggaact tttcgaaagg
     1681 tgaaagattg gttagacata cccgtactca taccggtgaa aaaccgtttc agtgtcggat
     1741 ctgtatgcga aatttctcca gaatggataa tttgtctact catctacgta cgcacaccgg
     1801 cgagaagcca ttccaatgcc gaatatgcat gcgcaacttc agtagaaaag atgctttgaa
     1861 tagacaccta aaaacccacc tgagaggatc cgggcgcgcc gacgcgctgg acgatttcga
     1921 tctcgacatg ctgggttctg atgccctcga tgactttgac ctggatatgt tgggaagcga
     1981 cgcattggat gactttgatc tggacatgct cggctccgat gctctggacg atttcgatct
     2041 cgatatgtta attaattgtt aagctagctg gccagacatg ataagataca ttgatgagtt
     2101 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2161 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2221 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2281 ctacaaatgt ggtatggaaa tgttaattaa ctagccatga ccaaaatccc ttaacgtgag
     2341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2521 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tcttaattaa cctgcaggcg
     3061 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3361 tgatgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttgactagtg gagccgagag
     3481 taattcatac aaaaggaggg atcgccttcg caaggggaga gcccagggac cgtccctaaa
     3541 ttctcacaga cccaaatccc tgtagccgcc ccacgacagc gcgaggagca tgcgcccagg
     3601 gctgagcgcg ggtagatcag agcacacaag ctcacagtcc ccggcggtgg ggggaggggc
     3661 gcgctgagcg ggggccaggg agctggcgcg gggcaaactg ggaaagtggt gtcgtgtgct
     3721 ggctccgccc tcttcccgag ggtgggggag aacggtatat aagtgcggta gtcgccttgg
     3781 acgttctttt tcgcaacggg tttgccgtca gaacgcaggt gagtggcggg tgtggcttcc
     3841 gcgggccccg gagctggagc cctgctctga gcgggccggg ctgatatgcg agtgtcgtcc
     3901 gcagggttta gctgtgagca ttcccacttc gagtggcggg cggtgcgggg gtgagagtgc
     3961 gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg cttgaggcct agcgtggtgt
     4021 ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt gtccttgctg ccctcgattg
     4081 ccttccagca gcatgggcta acaaagggag ggtgtggggc tcactcttaa ggagcccatg
     4141 aagcttacgt tggataggaa tggaagggca ggaggggcga ctggggcccg cccgccttcg
     4201 gagcacatgt ccgacgccac ctggatgggg cgaggcctgt ggctttccga agcaatcggg
     4261 cgtgagttta gcctacctgg gccatgtggc cctagcactg ggcacggtct ggcctggcgg
     4321 tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc cgcccggcac cagttgcttg
     4381 cgcggaaaga tggccgctcc cggggccctg ttgcaaggag ctcaaaatgg aggacgcggc
     4441 agcccggtgg agcgggcggg tgagtcaccc acacaaagga agagggcctt gcccctcgcc
     4501 ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat agtcacctcg ggcttctctt
     4561 gagcaccgct cgtcgcggcg gggggagggg atctaatggc gttggagttt gttcacattt
     4621 ggtgggtgga gactagtcag gccagcctgg cgctggaagt cattcttgga atttgcccct
     4681 ttgagtttgg agcgaggcta attctcaagc ctcttagcgg ttcaaaggta ttttctaaac
     4741 ccgtttccag gtgttgtgaa agccaccgct aattcaaagc aatccggagt atacatggtg
     4801 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     4861 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4921 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     4981 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     5041 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     5101 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     5161 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     5221 aagaagacca tgggctggca ggcctcctcc gagcggatgt accccgagga cggcgccctg
     5281 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     5341 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     5401 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     5461 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaacctag gagcaggttt
     5521 ccccaatgac acaaaacgtg caacttgaaa ctccgcctgg tctttccagg tctagagggg
     5581 taacactttg tactgcgttt ggctccacgc tcgatccact ggcgagtgtt agtaacagca
     5641 ctgttgcttc gtagcggagc atgacggccg tgggaactcc tccttggtaa caaggaccca
     5701 cggggccaaa agccacgccc acacgggccc gtcatgtgtg caaccccagc acggcgactt
     5761 tactgcgaaa cccactttaa agtgacattg aaactggtac ccacacactg gtgacaggct
     5821 aaggatgccc ttcaggtacc ccgaggtaac acgcgacact cgggatctga gaaggggact
     5881 ggggcttcta taaaagcgct cggtttaaaa agcttctatg cctgaatagg tgaccggagg
     5941 tcggcacctt tcctttgcaa ttactgaccc tatgaataca ctgactgttt gacaattaat
     6001 catcggcata gtatatcggc atagtataat acgactcact ataggagggc caccatgatt
     6061 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6121 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6181 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     6241 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6301 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6361 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6421 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6481 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6541 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     6601 gatctcgtcg tgacacatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6661 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6721 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6781 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6841 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgaattcg ctaggattat
     6901 ccctaatacc tgccacccca ctcttaatca gtggtggaag aacggtctca gaactgtttg
     6961 tttcaattgg ccatttaagt ttagtagtaa aagactggtt aatgataaca atgcatcgta
     7021 aaaccttcag aaggaaagga gaatgttttg tggaccactt tggttttctt ttttgcgtgt
     7081 ggcagtttta agttattagt ttttaaaatc agtacttttt aatggaaaca acttgaccaa
     7141 aaatttgtca cagaattttg agacccatta aaaaagttaa atgagaaacc tgtgtgttcc
     7201 tttggtcaac accgagacat ttaggtgaaa gacatctaat tctggtttta cgaatctgga
     7261 aacttcttga aaatgtaatt cttgagttaa cacttctggg tggagaatag ggttgttttc
     7321 cccccacata attggaaggg gaaggaatat catttaaagc tatgggaggg ttgctttgat
     7381 tacaacactg gagagaaatg cagcatgttg ctgattgcct gtcactaaaa caggccaaaa
     7441 actgagtcct tgggttgcat agaaagctg
//
