LOCUS       sequence-385007-        5564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient, constitutive expression of synZiFTR: ZF6-p65.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5564)
  AUTHORS   Li HS, Israni DV, Gagnon KA, Gan KA, Raymond MH, Sander JD, Roybal 
            KT, Joung JK, Wong WW, Khalil AS
  TITLE     Multidimensional control of therapeutic human cell function with 
            synthetic gene circuits.
  JOURNAL   Science. 2022 Dec 16;378(6625):1227-1234. doi: 
            10.1126/science.ade0156. Epub 2022 Dec 15.
  PUBMED    36520914
REFERENCE   2  (bases 1 to 5564)
  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"; doi: 
            "10.1126/science.ade0156"; journalName: "Science"; date: 
            "2022-12-16- 16"; volume: "378"; issue: "6625"; pages: "1227-1234"
COMMENT     Sequence Label: pMZ-181
FEATURES             Location/Qualifiers
     source          1..5564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(88..879,880..948))
                     /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     711..730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(949..1053)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1097..2308
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2145..2164
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      2326..2335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2335..2400
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2407..2427
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3226..3597
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     primer_bind     complement(3686..3705)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3751..3806
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3805..3824)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4167..4183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4167..4183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4180..4202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4191..4207
                     /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(4215..4221,4222..4239,4240..4245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4260..4281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      4366..4500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4520..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4557..4576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4611..4630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4722..4739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4893..5481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4973..4992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
       61 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      121 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      181 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      241 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      301 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
      361 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
      421 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      481 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
      541 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
      601 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
      661 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
      721 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
      781 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
      841 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
      901 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
      961 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1021 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     1081 gtcgacattg attattgact agtgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     1141 ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc
     1201 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     1261 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     1321 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     1381 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     1441 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     1501 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     1561 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     1621 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     1681 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     1741 aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat gggctggggc
     1801 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     1861 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     1921 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     1981 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     2041 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     2101 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     2161 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     2221 gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt
     2281 ctggccgttt ttggcttttt tgttagacac gcgtttaatt aagccgccac catggactac
     2341 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     2401 atggccccca agaaaaagag gaaggtgggc attcacgggg tgccgggtgg actcgaggga
     2461 ggcggtggaa gcggcggtac cgctagctct agaccaggcg aacgaccgtt tcaatgccgg
     2521 atatgtatga ggaacttctc ccagcagacc aatctcacgc ggcacacccg aacacataca
     2581 ggagaaaagc cattccaatg tcgaatttgt atgcgcaatt tttcacagaa ggctaacctc
     2641 ggcgttcacc tccgcacaca tactggttca cagaagccct ttcagtgcag gatttgtatg
     2701 cgcaacttta gcgatcactc atctctgaaa cgccacacta gaacgcatac aggtgagaag
     2761 cctttccagt gtcgcatctg tatgcggaac ttcagccgag cggatatgct tagacgccat
     2821 ttgcgaactc acaccggttc tcaaaaacct tttcagtgcc gaatttgtat gcgcaacttc
     2881 agcgagaaac agaacctggc acgccacacg cgcacgcaca ccggggaaaa accgtttcaa
     2941 tgtcgaatct gcatgcgcaa ttttagcgac ccctctaatc ttaggaggca tctgcgcaca
     3001 cacttgcggg gatccacctg cagggatgag tttcccacca tggtgtttcc ttctgggcag
     3061 atcagccagg cctcggcctt ggccccggcc cctccccaag tcctgcccca ggctccagcc
     3121 cctgcccctg ctccagccat ggtatcagct ctggcccagg ccccagcccc tgtcccagtc
     3181 ctagccccag gccctcctca ggctgtggcc ccacctgccc ccaagcccac ccaggctggg
     3241 gaaggaacgc tgtcagaggc cctgctgcag ctgcagtttg atgatgaaga cctgggggcc
     3301 ttgcttggca acagcacaga cccagctgtg ttcacagacc tggcatccgt cgacaactcc
     3361 gagtttcagc agctgctgaa ccagggcata cctgtggccc cccacacaac tgagcccatg
     3421 ctgatggagt accctgaggc tataactcgc ctagtgacag gggcccagag gccccccgac
     3481 ccagctcctg ctccactggg ggccccgggg ctccccaatg gcctcctttc aggagatgaa
     3541 gacttctcct ccattgcgga catggacttc tcagccctgc tgagtcagat cagctcctaa
     3601 gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     3661 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     3721 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     3781 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     3841 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     3901 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     3961 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     4021 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4081 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     4141 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4321 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     4381 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4441 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4501 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4621 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     4681 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatc
//
