LOCUS       sequence-270551-        7061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPI (BiFC/TriFC) assay vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7061)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 7061)
  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"
COMMENT     Sequence Label: pRDAX-518    N-Tag GFP[2] L10 Tln1(rod) EBPF2
FEATURES             Location/Qualifiers
     source          1..7061
                     /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(2220..2794,2795..2797,2798..2806)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2387..2404)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2614..2633
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2807..2809,2810..2812,2813..3526)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(2852..2873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3572..3704
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3713..4301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3766..3786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4184..4195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4328..4345)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4334..4445
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4541..4561)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4541..4559)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4580..4596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4580..4596)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4593..4615)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(4698..4717)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4763..4818
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4817..4836)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(5197..5785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5277..5296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5956..6747,6748..6816))
                     /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     6579..6598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6817..6921)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6989..7007
                     /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 caccatggaa aagagggacc acatggtgct gctcgagtac gtcacagcag ctggcatcac
     1081 cgatgccagc gggagctctg ccagcggtgg cagcacctcc gctgtgtctg gtatcattgc
     1141 tgacctcgac accaccatca tgttcgctac tgctggcaca cttaaccgtg agggtgctga
     1201 aacttttgct gaccaccggg agggtatctt aaagacagca aaggttcttg tggaggacac
     1261 caaggtccta gtgcagaatg cagctgggag ccaggagaag ttggcacaag ccgcccagtc
     1321 ctccgtggcc accattaccc gcctcgctga tgtggtcaag ctcggtgcag ccagcctagg
     1381 agccgaagac cctgaaactc aggtggtgct gatcaatgca gtaaaggacg tagccaaggc
     1441 cctgggtgac ctcatcagcg ctacgaaggc tgcagcgggc aaagttgggg atgaccctgc
     1501 agtgtggcag ctcaagaact ctgccaaggt gatggtgacc aatgtgacat cattgctcaa
     1561 gacagtgaag gctgtggaag atgaggccac caaaggcaca cgggccctag aggcaaccac
     1621 agagcacata cgtcaggaac tggcggtctt ctgttcccca gagccacctg ccaagacctc
     1681 tacccctgaa gacttcatcc gaatgaccaa gggtattact atggcaacag ccaaagccgt
     1741 tgctgctggc aattcctgtc gacaggaaga tgtcattgcc acagccaatc tgagccgacg
     1801 ggctattgcg gacatgcttc gggcttgcaa ggaagcagct ttccacccag aagtggcgcc
     1861 tgatgtacgg ctccgagccc tgcactatgg ccgggagtgt gccaatggtt acctggaact
     1921 gctggaccac gtgctgctga cccttcagaa gccaaaccca gacctgaagc agcagctaac
     1981 tggacactca aagcgggttg ctggctctgt gactgagctc atccaggctg ctgaagccat
     2041 gaaaggaaca gagtgggtgg acccagagga ccctactgtc attgctgaga atgagctctt
     2101 gggagccgca gccgccatcg aggctgcagc caagaagctg gagcagctga agcccagggc
     2161 caaacccaag gaggcggatg agtccttgaa ctttgaggaa caataagaat tcagatccgc
     2221 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     2281 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     2341 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     2401 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggtagcttc ttgaagacaa
     2461 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2521 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2581 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2641 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2701 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2761 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     2821 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2881 ccacaagttc agcgtgaggg gcgagggcga gggcgatgcc accaacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3001 gagccacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     3061 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3121 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3181 gctgaagggc gtcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3241 cttcaacagc cacaacatct atatcatggc cgtcaagcag aagaacggca tcaaggtgaa
     3301 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     3361 gaacaccccc atcggcgacg gccccgtgct gctgcccgac agccactacc tgagcaccca
     3421 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttccg
     3481 caccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaaccg gtgctcgagg
     3541 taccgatgaa tagctaaggt agaggccgca ggtaagtatc aaggttacaa gacaggttta
     3601 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     3661 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcg acaatcaacc
     3721 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3781 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3841 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3901 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3961 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4021 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4081 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     4141 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4201 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4261 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcggta
     4321 cgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4381 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4441 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4501 aagggggagg attgggaaga caatagccca gcttttgttc cctttagtga gggttaattg
     4561 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctaattc
     4621 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     4681 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     4741 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     4801 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     4861 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     4921 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     4981 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     5041 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     5101 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatcggccgg ccgccggcgt
     5161 cgacgcgcgc ggccgcggcg cgccgttgct ggcgtttttc cataggctcc gcccccctga
     5221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6001 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6061 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6121 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6181 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6241 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6301 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6361 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6421 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6481 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6541 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6601 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6661 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6721 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6781 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6841 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6901 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6961 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctacg
     7021 cgcgcgccgg cggcgcgccg tcgacggccg gccgcggccg c
//
