LOCUS       sequence-343591-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive PhCMV-driven mammalian expression vector of CDH-1-TF 
            cassette (PhCMV-Gal4-Bcl-XL-P2A-LD3-Rel65-pA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  AUTHORS   Shui S, Gainza P, Scheller L, Yang C, Kurumida Y, Rosset S, Georgeon
            S, Di Roberto RB, Castellanos-Rueda R, Reddy ST, Correia BE
  TITLE     A rational blueprint for the design of chemically-controlled protein
            switches.
  JOURNAL   Nat Commun. 2021 Oct 1;12(1):5754. doi: 10.1038/s41467-021-25735-9.
  PUBMED    34599176
REFERENCE   2  (bases 1 to 7283)
  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 
            Commun."; date: "2021-10-1"; volume: "12(1):5754. doi"; pages: " 
            10.1038/s41467-021-25735-9"
COMMENT     Sequence Label: S111
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             895..1335
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1867..1923
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1939..1956
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2416..2787
                     /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"
     CDS             2797..2817
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2836..2854)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2837..2854)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2874..2891)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2880..3104
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3150..3578
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3237..3256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3447..3468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3587..3607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3592..3921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3772..3907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3834..3853
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3988..4782
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4042..4061)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4652..4671
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4956..5077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4993..5012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5047..5066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5126..5142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5126..5142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5139..5161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5150..5166
                     /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(5174..5180,5181..5198,5199..5204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5219..5240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5357..5374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5528..6116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5608..5627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6287..7078,7079..7147))
                     /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     6910..6929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7148..7252)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgaag
      901 ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct caagtgctcc
      961 aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg ctactctccc
     1021 aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc aaggctagaa
     1081 agactggaac agctatttct actgattttt cctcgagaag accttgacat gattttgaaa
     1141 atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca agataatgtg
     1201 aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc tctaacattg
     1261 agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa aggtcaaaga
     1321 cagttgactg tatcggaatt catgagccag agcaaccgtg agctggttgt ggacttcctg
     1381 agctacaagc tgagccagaa aggttatagc tggagccaat ttagcgatgt ggaggaaaac
     1441 cgtaccgagg cgccggaagg taccgagagc gaagcggtta agcaggcgct gcgtgaagcg
     1501 ggcgacgagt tcgaactgcg ttaccgtcgt gcgtttagcg atctgaccag ccaactgcac
     1561 atcaccccgg gcaccgcgta tcagagcttc gagcaagtgg ttaacgaact gtttcgtgac
     1621 ggtgtgaact ggggccgtat tgttgcgttc tttagctttg gtggcgcgct gtgcgtggag
     1681 agcgttgata aggaaatgca ggtgctggtt agccgtatcg cggcgtggat ggcgacctac
     1741 ctgaacgacc acctggagcc gtggattcaa gaaaacggtg gctgggatac ctttgtggag
     1801 ctgtatggta acaacgcggc ggcggagagc cgtaaaggcc aagaacgtga ggataatccc
     1861 tgcagggcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
     1921 ccgcctgcag ggactatgca tcaccatcac catcacggca gcggtcagcg ttgggaactg
     1981 gcactgggtc gctttctgga atatctgagt tgggtctcca ccctgtcaga acaggtgcaa
     2041 gaagaactgc tgtcttctca ggtgacccaa gaactgcgtg cgctgatgga tgaaacgatg
     2101 aaagaactga aagcctacaa atctgaactg gaagaacagc tgaccccggt tgcagaagaa
     2161 acgcgtgctc gcctgtccaa agaactgcag gccgcacaag cccgtctggg tgcagatatg
     2221 gaagacgtgc gtggccgcct ggttcagtat cgcggtgaag ttcaggccat gctgggccaa
     2281 tcgacggaag aactgcgtgt ccgcctggcg agccatctga ttgccctgca gctgcgtctg
     2341 atcggtgatg cattcgacct gcagaaacgc ctggctgtct accaagcagg cgctgctgag
     2401 gctagcggtg gaggacccac ccaggctggg gaaggaacgc tgtcagaggc cctgctgcag
     2461 ctgcagtttg atgatgaaga cctgggggcc ttgcttggca acagcacaga cccagctgtg
     2521 ttcacagacc tggcatccgt cgacaactcc gagtttcagc agctgctgaa ccagggcata
     2581 cctgtggccc cccacacaac tgagcccatg ctgatggagt accctgaggc tataactcgc
     2641 ctagtgacag gggcccagag gccccccgac ccagctcctg ctccactggg ggccccgggg
     2701 ctccccaatg gcctcctttc aggagatgaa gacttctcct ccattgcgga catggacttc
     2761 tcagccctgc tgagtcagat cagctccctg tacaagccaa agaagaagag aaaggtataa
     2821 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2881 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2941 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3001 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3061 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3121 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3181 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3241 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3301 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3361 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3421 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3481 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3541 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3601 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3661 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3781 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3841 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3901 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3961 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4021 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4081 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4141 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4201 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4261 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4321 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4381 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4441 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4501 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4561 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4621 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4681 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4741 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4801 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4861 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4921 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4981 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5041 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5101 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5161 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5221 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5281 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5341 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7261 cccgaaaagt gccacctgac gtc
//
