LOCUS       sequence-120992-        7733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF8/ZF7-TF with mCherry marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7733)
  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 7733)
  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-148
FEATURES             Location/Qualifiers
     source          1..7733
                     /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             2165..2314
                     /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    2346..2467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2357..2376)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2411..2430
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2655..3243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3144..3163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3323..3626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3741..5046
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    4083..5014
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             5059..5769
                     /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(5204..5222)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5472..5491
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5776..6220
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6253..6299
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6291..6309
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6319..7113
                     /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             6319..7113
                     /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(6373..6392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6983..7002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7160..7732
                     /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 tttcgactat
     1681 ggctgttttg agaagacata cccgtactca taccggtgaa aaaccgtttc agtgtcggat
     1741 ctgtatgcga aatttctcca gaagagaagt tttggaaaat catctacgta cgcacaccgg
     1801 cgagaagcca ttccaatgcc gaatatgcat gcgcaacttc agtcaaactg ttaatttgga
     1861 tagacaccta aaaacccacc tgagacagaa ggacggggag cgccccttcc agtgtcgcat
     1921 ttgcatgcgg aacttttcga aaaaagatca tttgcataga catacccgta ctcataccgg
     1981 tgaaaaaccg tttcagtgtc ggatctgtat gcgaaatttc tcccaaagac cacatttgac
     2041 taatcatcta cgtacgcaca ccggcgagaa gccattccaa tgccgaatat gcatgcgcaa
     2101 cttcagtgtt ggtgcttctt tgaaaagaca cctaaaaacc cacctgagag gatccgggcg
     2161 cgccgacgcg ctggacgatt tcgatctcga catgctgggt tctgatgccc tcgatgactt
     2221 tgacctggat atgttgggaa gcgacgcatt ggatgacttt gatctggaca tgctcggctc
     2281 cgatgctctg gacgatttcg atctcgatat gttaattaat tgttaagcta gctggccaga
     2341 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2401 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2461 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2521 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaaatgttaa ttaactagcc
     2581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2761 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3301 atgttcttaa ttaacctgca ggcgttacat aacttacggt aaatggcccg cctggctgac
     3361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     3601 acgtattagt catcgctatt accatgatga tgcggttttg gcagtacatc aatgggcgtg
     3661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     3721 tgttttgact agtggagccg agagtaattc atacaaaagg agggatcgcc ttcgcaaggg
     3781 gagagcccag ggaccgtccc taaattctca cagacccaaa tccctgtagc cgccccacga
     3841 cagcgcgagg agcatgcgcc cagggctgag cgcgggtaga tcagagcaca caagctcaca
     3901 gtccccggcg gtggggggag gggcgcgctg agcgggggcc agggagctgg cgcggggcaa
     3961 actgggaaag tggtgtcgtg tgctggctcc gccctcttcc cgagggtggg ggagaacggt
     4021 atataagtgc ggtagtcgcc ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc
     4081 aggtgagtgg cgggtgtggc ttccgcgggc cccggagctg gagccctgct ctgagcgggc
     4141 cgggctgata tgcgagtgtc gtccgcaggg tttagctgtg agcattccca cttcgagtgg
     4201 cgggcggtgc gggggtgaga gtgcgaggcc tagcggcaac cccgtagcct cgcctcgtgt
     4261 ccggcttgag gcctagcgtg gtgtccgccg ccgcgtgcca ctccggccgc actatgcgtt
     4321 ttttgtcctt gctgccctcg attgccttcc agcagcatgg gctaacaaag ggagggtgtg
     4381 gggctcactc ttaaggagcc catgaagctt acgttggata ggaatggaag ggcaggaggg
     4441 gcgactgggg cccgcccgcc ttcggagcac atgtccgacg ccacctggat ggggcgaggc
     4501 ctgtggcttt ccgaagcaat cgggcgtgag tttagcctac ctgggccatg tggccctagc
     4561 actgggcacg gtctggcctg gcggtgccgc gttcccttgc ctcccaacaa gggtgaggcc
     4621 gtcccgcccg gcaccagttg cttgcgcgga aagatggccg ctcccggggc cctgttgcaa
     4681 ggagctcaaa atggaggacg cggcagcccg gtggagcggg cgggtgagtc acccacacaa
     4741 aggaagaggg ccttgcccct cgccggccgc tgcttcctgt gaccccgtgg tctatcggcc
     4801 gcatagtcac ctcgggcttc tcttgagcac cgctcgtcgc ggcgggggga ggggatctaa
     4861 tggcgttgga gtttgttcac atttggtggg tggagactag tcaggccagc ctggcgctgg
     4921 aagtcattct tggaatttgc ccctttgagt ttggagcgag gctaattctc aagcctctta
     4981 gcggttcaaa ggtattttct aaacccgttt ccaggtgttg tgaaagccac cgctaattca
     5041 aagcaatccg gagtatacat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     5101 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     5161 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     5221 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     5281 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     5341 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     5401 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     5461 ccctccgacg gccccgtaat gcagaagaag accatgggct ggcaggcctc ctccgagcgg
     5521 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     5581 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     5641 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     5701 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     5761 tacaagtaac ctaggagcag gtttccccaa tgacacaaaa cgtgcaactt gaaactccgc
     5821 ctggtctttc caggtctaga ggggtaacac tttgtactgc gtttggctcc acgctcgatc
     5881 cactggcgag tgttagtaac agcactgttg cttcgtagcg gagcatgacg gccgtgggaa
     5941 ctcctccttg gtaacaagga cccacggggc caaaagccac gcccacacgg gcccgtcatg
     6001 tgtgcaaccc cagcacggcg actttactgc gaaacccact ttaaagtgac attgaaactg
     6061 gtacccacac actggtgaca ggctaaggat gcccttcagg taccccgagg taacacgcga
     6121 cactcgggat ctgagaaggg gactggggct tctataaaag cgctcggttt aaaaagcttc
     6181 tatgcctgaa taggtgaccg gaggtcggca cctttccttt gcaattactg accctatgaa
     6241 tacactgact gtttgacaat taatcatcgg catagtatat cggcatagta taatacgact
     6301 cactatagga gggccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6361 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6421 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6481 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     6541 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6601 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6661 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6721 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6781 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6841 aaggcgagca tgcccgacgg cgaggatctc gtcgtgacac atggcgatgc ctgcttgccg
     6901 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6961 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7021 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7081 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7141 accaagcgaa ttcgctagga ttatccctaa tacctgccac cccactctta atcagtggtg
     7201 gaagaacggt ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact
     7261 ggttaatgat aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc
     7321 actttggttt tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact
     7381 ttttaatgga aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag
     7441 ttaaatgaga aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc
     7501 taattctggt tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc
     7561 tgggtggaga atagggttgt tttcccccca cataattgga aggggaagga atatcattta
     7621 aagctatggg agggttgctt tgattacaac actggagaga aatgcagcat gttgctgatt
     7681 gcctgtcact aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctg
//
