LOCUS       sequence-249239-        6213 bp    DNA     circular SYN 15-APR-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 6213)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRDAX-358 N-Tag Venus173/155[2] L20 {Esp3I(2x)...
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1019..1028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(1381..1955,1956..1958,1959..1967)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1548..1565)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1775..1794
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1968..2678
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2113..2131)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2381..2400
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2724..2856
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2865..3453
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2918..2938)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3336..3347)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3480..3497)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3486..3597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3693..3713)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3693..3711)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3732..3748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3732..3748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3745..3767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3850..3869)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3915..3970
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3969..3988)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4349..4937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4429..4448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5108..5899,5900..5968))
                     /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     5731..5750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5969..6073)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6141..6159
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcctgcctcc aggcctcaat attggccatt agccatatta ttcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag
      841 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccaggc
     1021 caccatggat aagcagaaga atgggattaa ggcgaacttc aagatccgac acaacatcga
     1081 ggatggtggg gtccagcttg ccgaccatta tcagcagaac acacccattg gggatggacc
     1141 agtcctgttg ccggataacc actacctcag ttaccagagc aaactgtcca aagatcccaa
     1201 tgagaagagg gaccatatgg tactgctcga attcgtgact gcagcaggca ttacccttgg
     1261 gatggatgaa ctttataagg ggagtcctgg cggtggatcc ggaggctctg ggagctctgc
     1321 cagcggtggc agcacctccc gagacgccga ggcacgccgt ctcctaagaa ttcagatccg
     1381 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     1441 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     1501 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     1561 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggtagctt cttgaagaca
     1621 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     1681 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     1741 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     1801 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggta
     1861 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     1921 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     1981 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     2041 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     2101 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     2161 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     2221 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     2281 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     2341 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2401 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     2461 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     2521 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     2581 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     2641 gccactccac cggcggcatg gacgagctgt acaagtaaac cggtgctcga ggtaccgatg
     2701 aatagctaag gtcgaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc
     2761 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     2821 gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa cctctggatt
     2881 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2941 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3001 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3061 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3121 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3181 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3241 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3301 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3361 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3421 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacgatcagc
     3481 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3601 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3661 ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3721 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctaat tcactcctca
     3781 ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     3841 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     3901 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3961 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     4021 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     4081 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     4141 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     4201 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     4261 cccagtcata gctgtccctc ttctcttatg gagatcggcc ggccgccggc gtcgacgcgc
     4321 gcggccgcgg cgcgccgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5161 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5221 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5281 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5341 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5401 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5461 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5521 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5581 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5641 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5701 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5761 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5821 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5881 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6001 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6061 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6121 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtcta cgcgcgcgcc
     6181 ggcggcgcgc cgtcgacggc cggccgcggc cgc
//
