LOCUS       sequence-121009-        7525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF9-TF-V2 with mCherry marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7525)
  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 7525)
  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-247
FEATURES             Location/Qualifiers
     source          1..7525
                     /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             1585..1608
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1609..1629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1957..2106
                     /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    2138..2259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2149..2168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2203..2222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2447..3035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2936..2955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3115..3418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3533..4838
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3875..4806
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             4851..5561
                     /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(4996..5014)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5264..5283
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5568..6012
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6045..6091
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6083..6101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6111..6905
                     /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             6111..6905
                     /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(6165..6184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6775..6794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6952..7524
                     /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 atcaaagatc tatggactac aaggacgacg atgacaagcc caagaaaaag
     1621 cgcaaggtct ctagacccgg ggagcgcccc ttccagtgtc gcatttgcat gcggaacttt
     1681 tcggataaaa ctaaattgag agttcatacc cgtactcata ccggtgaaaa accgtttcag
     1741 tgtcggatct gtatgcgaaa tttctccgtt agacataatt tgactagaca tctacgtacg
     1801 cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag tcaatctact
     1861 tctttgcaaa gacacctaaa aacccacctg agaggatccg gcggagtgct ggagaagggc
     1921 tgcaccatga ccgagaaggg cagcggaggg cgcgccgacg cgctggacga tttcgatctc
     1981 gacatgctgg gttctgatgc cctcgatgac tttgacctgg atatgttggg aagcgacgca
     2041 ttggatgact ttgatctgga catgctcggc tccgatgctc tggacgattt cgatctcgat
     2101 atgttaatta attgttaagc tagctggcca gacatgataa gatacattga tgagtttgga
     2161 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2221 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2281 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2341 aaatgtggta tggaaatgtt aattaactag ccatgaccaa aatcccttaa cgtgagtttt
     2401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3061 ggttcctggc cttttgctgg ccttttgctc acatgttctt aattaacctg caggcgttac
     3121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgat
     3421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3481 aagtctccac cccattgacg tcaatgggag tttgttttga ctagtggagc cgagagtaat
     3541 tcatacaaaa ggagggatcg ccttcgcaag gggagagccc agggaccgtc cctaaattct
     3601 cacagaccca aatccctgta gccgccccac gacagcgcga ggagcatgcg cccagggctg
     3661 agcgcgggta gatcagagca cacaagctca cagtccccgg cggtgggggg aggggcgcgc
     3721 tgagcggggg ccagggagct ggcgcggggc aaactgggaa agtggtgtcg tgtgctggct
     3781 ccgccctctt cccgagggtg ggggagaacg gtatataagt gcggtagtcg ccttggacgt
     3841 tctttttcgc aacgggtttg ccgtcagaac gcaggtgagt ggcgggtgtg gcttccgcgg
     3901 gccccggagc tggagccctg ctctgagcgg gccgggctga tatgcgagtg tcgtccgcag
     3961 ggtttagctg tgagcattcc cacttcgagt ggcgggcggt gcgggggtga gagtgcgagg
     4021 cctagcggca accccgtagc ctcgcctcgt gtccggcttg aggcctagcg tggtgtccgc
     4081 cgccgcgtgc cactccggcc gcactatgcg ttttttgtcc ttgctgccct cgattgcctt
     4141 ccagcagcat gggctaacaa agggagggtg tggggctcac tcttaaggag cccatgaagc
     4201 ttacgttgga taggaatgga agggcaggag gggcgactgg ggcccgcccg ccttcggagc
     4261 acatgtccga cgccacctgg atggggcgag gcctgtggct ttccgaagca atcgggcgtg
     4321 agtttagcct acctgggcca tgtggcccta gcactgggca cggtctggcc tggcggtgcc
     4381 gcgttccctt gcctcccaac aagggtgagg ccgtcccgcc cggcaccagt tgcttgcgcg
     4441 gaaagatggc cgctcccggg gccctgttgc aaggagctca aaatggagga cgcggcagcc
     4501 cggtggagcg ggcgggtgag tcacccacac aaaggaagag ggccttgccc ctcgccggcc
     4561 gctgcttcct gtgaccccgt ggtctatcgg ccgcatagtc acctcgggct tctcttgagc
     4621 accgctcgtc gcggcggggg gaggggatct aatggcgttg gagtttgttc acatttggtg
     4681 ggtggagact agtcaggcca gcctggcgct ggaagtcatt cttggaattt gcccctttga
     4741 gtttggagcg aggctaattc tcaagcctct tagcggttca aaggtatttt ctaaacccgt
     4801 ttccaggtgt tgtgaaagcc accgctaatt caaagcaatc cggagtatac atggtgagca
     4861 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     4921 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     4981 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     5041 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     5101 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     5161 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     5221 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     5281 agaccatggg ctggcaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     5341 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     5401 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     5461 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     5521 gccgccactc caccggcggc atggacgagc tgtacaagta acctaggagc aggtttcccc
     5581 aatgacacaa aacgtgcaac ttgaaactcc gcctggtctt tccaggtcta gaggggtaac
     5641 actttgtact gcgtttggct ccacgctcga tccactggcg agtgttagta acagcactgt
     5701 tgcttcgtag cggagcatga cggccgtggg aactcctcct tggtaacaag gacccacggg
     5761 gccaaaagcc acgcccacac gggcccgtca tgtgtgcaac cccagcacgg cgactttact
     5821 gcgaaaccca ctttaaagtg acattgaaac tggtacccac acactggtga caggctaagg
     5881 atgcccttca ggtaccccga ggtaacacgc gacactcggg atctgagaag gggactgggg
     5941 cttctataaa agcgctcggt ttaaaaagct tctatgcctg aataggtgac cggaggtcgg
     6001 cacctttcct ttgcaattac tgaccctatg aatacactga ctgtttgaca attaatcatc
     6061 ggcatagtat atcggcatag tataatacga ctcactatag gagggccacc atgattgaac
     6121 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     6181 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     6241 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     6301 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6361 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6421 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6481 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6541 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6601 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     6661 tcgtcgtgac acatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6721 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6781 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6841 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6901 tctgagcggg actctggggt tcgaaatgac cgaccaagcg aattcgctag gattatccct
     6961 aatacctgcc accccactct taatcagtgg tggaagaacg gtctcagaac tgtttgtttc
     7021 aattggccat ttaagtttag tagtaaaaga ctggttaatg ataacaatgc atcgtaaaac
     7081 cttcagaagg aaaggagaat gttttgtgga ccactttggt tttctttttt gcgtgtggca
     7141 gttttaagtt attagttttt aaaatcagta ctttttaatg gaaacaactt gaccaaaaat
     7201 ttgtcacaga attttgagac ccattaaaaa agttaaatga gaaacctgtg tgttcctttg
     7261 gtcaacaccg agacatttag gtgaaagaca tctaattctg gttttacgaa tctggaaact
     7321 tcttgaaaat gtaattcttg agttaacact tctgggtgga gaatagggtt gttttccccc
     7381 cacataattg gaaggggaag gaatatcatt taaagctatg ggagggttgc tttgattaca
     7441 acactggaga gaaatgcagc atgttgctga ttgcctgtca ctaaaacagg ccaaaaactg
     7501 agtccttggg ttgcatagaa agctg
//
