LOCUS       sequence-343592-        7262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive PhCMV-driven mammalian expression vector of CDH-2-TF 
            cassette (PhCMV-Gal4-Bcl2-P2A-LD3-Rel65-pA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7262)
  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 7262)
  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: S112
FEATURES             Location/Qualifiers
     source          1..7262
                     /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             1846..1902
                     /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             1918..1935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2395..2766
                     /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             2776..2796
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2815..2833)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2816..2833)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2853..2870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2859..3083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3129..3557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3216..3235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3426..3447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3566..3586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3571..3900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3813..3832
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3967..4761
                     /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(4021..4040)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4631..4650
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4935..5056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4972..4991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5026..5045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5105..5121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5105..5121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5118..5140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5129..5145
                     /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(5153..5159,5160..5177,5178..5183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5198..5219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5336..5353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5507..6095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5587..5606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6266..7057,7058..7126))
                     /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     6889..6908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7127..7231)
                     /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 tatcggaagt catggcgcac gctgggagaa cagggtacga taaccgggag
     1381 atagtgatga agtacatcca ttataagctg tcgcagaggg gctacgagtg ggatgcggga
     1441 gatgatgtgg aagagaacag gactgaggcc ccagaaggga ctgaatcgga ggtggtccat
     1501 ctgaccctcc gccaggccgg cgacgacttc tcccgccgct accgccgcga cttcgccgag
     1561 atgtccagcc agctgcacct gacgcccttc accgcgcggg gacgctttgc cacggtggtg
     1621 gaggagctct tcagggacgg ggtgaactgg gggaggattg tggccttctt tgagttcggt
     1681 ggggtcatgt gtgtggagag cgtcaaccgg gagatgtcgc ccctggtgga caacatcgcc
     1741 ctgtggatga ctgagtacct gaaccggcac ctgcacacct ggatccagga taacggaggc
     1801 tgggatgcct ttgtggaact gtacggcccc agcatgcggt gcagggcaac aaacttctct
     1861 ctgctgaaac aagccggaga tgtcgaagag aatcctggac cgcctgcagg gactatgcat
     1921 caccatcacc atcacggcag cggtcagcgt tgggaactgg cactgggtcg ctttctggaa
     1981 tatctgagtt gggtctccac cctgtcagaa caggtgcaag aagaactgct gtcttctcag
     2041 gtgacccaag aactgcgtgc gctgatggat gaaacgatga aagaactgaa agcctacaaa
     2101 tctgaactgg aagaacagct gaccccggtt gcagaagaaa cgcgtgctcg cctgtccaaa
     2161 gaactgcagg ccgcacaagc ccgtctgggt gcagatatgg aagacgtgcg tggccgcctg
     2221 gttcagtatc gcggtgaagt tcaggccatg ctgggccaat cgacggaaga actgcgtgtc
     2281 cgcctggcga gccatctgat tgccctgcag ctgcgtctga tcggtgatgc attcgacctg
     2341 cagaaacgcc tggctgtcta ccaagcaggc gctgctgagg ctagcggtgg aggacccacc
     2401 caggctgggg aaggaacgct gtcagaggcc ctgctgcagc tgcagtttga tgatgaagac
     2461 ctgggggcct tgcttggcaa cagcacagac ccagctgtgt tcacagacct ggcatccgtc
     2521 gacaactccg agtttcagca gctgctgaac cagggcatac ctgtggcccc ccacacaact
     2581 gagcccatgc tgatggagta ccctgaggct ataactcgcc tagtgacagg ggcccagagg
     2641 ccccccgacc cagctcctgc tccactgggg gccccggggc tccccaatgg cctcctttca
     2701 ggagatgaag acttctcctc cattgcggac atggacttct cagccctgct gagtcagatc
     2761 agctccctgt acaagccaaa gaagaagaga aaggtataat ctagagggcc ctattctata
     2821 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2881 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2941 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3001 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3061 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3121 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3181 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3241 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3301 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3361 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3421 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3481 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3541 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3961 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4021 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4081 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4141 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4201 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4261 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4321 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4381 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4441 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4501 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4561 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4621 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4681 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4741 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4801 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4861 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4921 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4981 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5041 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5101 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5161 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5221 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5281 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5341 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5401 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6601 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6661 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6721 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6781 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6841 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6901 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6961 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7021 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7081 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7141 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7201 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7261 tc
//
