LOCUS       sequence-120964-        7477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ZF4-TF with mCherry marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  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 7477)
  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-117
FEATURES             Location/Qualifiers
     source          1..7477
                     /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             1909..2058
                     /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    2090..2211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2101..2120)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2155..2174
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2399..2987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2888..2907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3067..3370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3485..4790
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3827..4758
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             4803..5513
                     /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(4948..4966)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5216..5235
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5520..5964
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5997..6043
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6035..6053
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6063..6857
                     /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             6063..6857
                     /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(6117..6136)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6727..6746
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6904..7476
                     /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 tcgactgctg ctgttttgac tagacatacc cgtactcata ccggtgaaaa accgtttcag
     1741 tgtcggatct gtatgcgaaa tttctccgat agagctaatt tgactagaca tctacgtacg
     1801 cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag tagaattgat
     1861 aaattgggtg atcacctaaa aacccacctg agaggatccg ggcgcgccga cgcgctggac
     1921 gatttcgatc tcgacatgct gggttctgat gccctcgatg actttgacct ggatatgttg
     1981 ggaagcgacg cattggatga ctttgatctg gacatgctcg gctccgatgc tctggacgat
     2041 ttcgatctcg atatgttaat taattgttaa gctagctggc cagacatgat aagatacatt
     2101 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2161 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2221 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2281 taaaacctct acaaatgtgg tatggaaatg ttaattaact agccatgacc aaaatccctt
     2341 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2521 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc ttaattaacc
     3061 tgcaggcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3361 tattaccatg atgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     3421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt gactagtgga
     3481 gccgagagta attcatacaa aaggagggat cgccttcgca aggggagagc ccagggaccg
     3541 tccctaaatt ctcacagacc caaatccctg tagccgcccc acgacagcgc gaggagcatg
     3601 cgcccagggc tgagcgcggg tagatcagag cacacaagct cacagtcccc ggcggtgggg
     3661 ggaggggcgc gctgagcggg ggccagggag ctggcgcggg gcaaactggg aaagtggtgt
     3721 cgtgtgctgg ctccgccctc ttcccgaggg tgggggagaa cggtatataa gtgcggtagt
     3781 cgccttggac gttctttttc gcaacgggtt tgccgtcaga acgcaggtga gtggcgggtg
     3841 tggcttccgc gggccccgga gctggagccc tgctctgagc gggccgggct gatatgcgag
     3901 tgtcgtccgc agggtttagc tgtgagcatt cccacttcga gtggcgggcg gtgcgggggt
     3961 gagagtgcga ggcctagcgg caaccccgta gcctcgcctc gtgtccggct tgaggcctag
     4021 cgtggtgtcc gccgccgcgt gccactccgg ccgcactatg cgttttttgt ccttgctgcc
     4081 ctcgattgcc ttccagcagc atgggctaac aaagggaggg tgtggggctc actcttaagg
     4141 agcccatgaa gcttacgttg gataggaatg gaagggcagg aggggcgact ggggcccgcc
     4201 cgccttcgga gcacatgtcc gacgccacct ggatggggcg aggcctgtgg ctttccgaag
     4261 caatcgggcg tgagtttagc ctacctgggc catgtggccc tagcactggg cacggtctgg
     4321 cctggcggtg ccgcgttccc ttgcctccca acaagggtga ggccgtcccg cccggcacca
     4381 gttgcttgcg cggaaagatg gccgctcccg gggccctgtt gcaaggagct caaaatggag
     4441 gacgcggcag cccggtggag cgggcgggtg agtcacccac acaaaggaag agggccttgc
     4501 ccctcgccgg ccgctgcttc ctgtgacccc gtggtctatc ggccgcatag tcacctcggg
     4561 cttctcttga gcaccgctcg tcgcggcggg gggaggggat ctaatggcgt tggagtttgt
     4621 tcacatttgg tgggtggaga ctagtcaggc cagcctggcg ctggaagtca ttcttggaat
     4681 ttgccccttt gagtttggag cgaggctaat tctcaagcct cttagcggtt caaaggtatt
     4741 ttctaaaccc gtttccaggt gttgtgaaag ccaccgctaa ttcaaagcaa tccggagtat
     4801 acatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     4861 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     4921 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4981 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     5041 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     5101 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     5161 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     5221 taatgcagaa gaagaccatg ggctggcagg cctcctccga gcggatgtac cccgaggacg
     5281 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     5341 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     5401 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     5461 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taacctagga
     5521 gcaggtttcc ccaatgacac aaaacgtgca acttgaaact ccgcctggtc tttccaggtc
     5581 tagaggggta acactttgta ctgcgtttgg ctccacgctc gatccactgg cgagtgttag
     5641 taacagcact gttgcttcgt agcggagcat gacggccgtg ggaactcctc cttggtaaca
     5701 aggacccacg gggccaaaag ccacgcccac acgggcccgt catgtgtgca accccagcac
     5761 ggcgacttta ctgcgaaacc cactttaaag tgacattgaa actggtaccc acacactggt
     5821 gacaggctaa ggatgccctt caggtacccc gaggtaacac gcgacactcg ggatctgaga
     5881 aggggactgg ggcttctata aaagcgctcg gtttaaaaag cttctatgcc tgaataggtg
     5941 accggaggtc ggcacctttc ctttgcaatt actgacccta tgaatacact gactgtttga
     6001 caattaatca tcggcatagt atatcggcat agtataatac gactcactat aggagggcca
     6061 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6121 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6181 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6241 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6301 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6361 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6421 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6481 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6541 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     6601 acggcgagga tctcgtcgtg acacatggcg atgcctgctt gccgaatatc atggtggaaa
     6661 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6721 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6781 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6841 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgaattcgct
     6901 aggattatcc ctaatacctg ccaccccact cttaatcagt ggtggaagaa cggtctcaga
     6961 actgtttgtt tcaattggcc atttaagttt agtagtaaaa gactggttaa tgataacaat
     7021 gcatcgtaaa accttcagaa ggaaaggaga atgttttgtg gaccactttg gttttctttt
     7081 ttgcgtgtgg cagttttaag ttattagttt ttaaaatcag tactttttaa tggaaacaac
     7141 ttgaccaaaa atttgtcaca gaattttgag acccattaaa aaagttaaat gagaaacctg
     7201 tgtgttcctt tggtcaacac cgagacattt aggtgaaaga catctaattc tggttttacg
     7261 aatctggaaa cttcttgaaa atgtaattct tgagttaaca cttctgggtg gagaataggg
     7321 ttgttttccc cccacataat tggaagggga aggaatatca tttaaagcta tgggagggtt
     7381 gctttgatta caacactgga gagaaatgca gcatgttgct gattgcctgt cactaaaaca
     7441 ggccaaaaac tgagtccttg ggttgcatag aaagctg
//
