LOCUS       sequence-249390-        8999 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 8999)
  TITLE     Lewin Lab Plasmids
REFERENCE   2  (bases 1 to 8999)
  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-533    C-Tag Venus210[1] L32 Itgb1 EBPF2
FEATURES             Location/Qualifiers
     source          1..8999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          871..1003
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    join(4157..4731,4732..4734,4735..4743)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4324..4341)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4551..4570
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4744..4746,4747..4749,4750..5463)
                     /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(4789..4810)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          5509..5641
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5650..6238
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5703..5723)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6121..6132)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6265..6282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6271..6382
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6478..6498)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6478..6496)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6517..6533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6517..6533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6530..6552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(6635..6654)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6700..6755
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6754..6773)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(7134..7722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7214..7233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7893..8684,8685..8753))
                     /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     8516..8535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8754..8858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8926..8944
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctgcctcca ggcctcaata ttggccatta gccatattat tcattggtta tatagcataa
       61 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
      121 attggctcat gtccaatatg accgccatgt tggcattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
      901 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
      961 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggcc
     1021 accatgaatt tgcaactggt ttcctggatt ggattgatca gtttgatttg ttctgtattt
     1081 ggccaaacag ataaaaatag atgtttaaaa gcaaatgcca aatcttgcgg agaatgtata
     1141 caagcagggc caaattgtgg gtggtgtaca aatacgacat ttttgcaaga aggaatgcct
     1201 acttctgcac gatgtgatga tttagaagct ttgaaaaaga agggttgcca gccaagtgac
     1261 atagagaatc ccagaggctc tcaaactata aagaaaaata aaaatgtcac caatcgcagc
     1321 aaagggatgg cagagaagct ccggccagaa gacattactc agatccaacc acaacagctg
     1381 cttctaaaat tgagatcagg agaaccacag aagtttacat taaaattcaa gagggctgaa
     1441 gattacccta ttgatctcta ctaccttatg gatctctcct actctatgaa agatgatctg
     1501 gagaatgtga aaagtcttgg aacggatttg atgaatgaaa tgaggaggat tacttcagac
     1561 ttccgcattg gctttggctc atttgtggag aaaactgtga tgccgtatat tagcacaacc
     1621 ccagcaaagc taagaaatcc ttgtacaagt gaacaaaact gcaccagccc atttagctac
     1681 aaaaatgtgc ttagtcttac tgacagagga gagtttttca atgaacttgt tggtcagcaa
     1741 cgcatatctg gaaacttgga ttctccagaa ggtggctttg atgcaatcat gcaggttgcg
     1801 gtttgtggat cgctgattgg ctggaggaat gtaacacgac tgctggtgtt ttccacggat
     1861 gctgggtttc actttgctgg agatgggaaa cttggtggta ttgttttacc caatgatgga
     1921 caatgtcacc tggaaaataa tgtatataca atgagccatt actatgatta tccttcaatt
     1981 gctcaccttg ttcagaaact aagtgaaaat aatattcaga cgatttttgc agttactgaa
     2041 gagttccaac ctgtttacaa ggaattgaag aatttgattc ctaagtcagc agtgggcaca
     2101 ctgtctggaa actctagtaa tgtgatccag ctaatcatcg atgcctacaa ctctctttct
     2161 tcagaagtca ttctggaaaa tagcaaattg ccagacggag taacaataaa ttacaaatcc
     2221 tattgcaaga atggggtgaa tgggacagga gaaaatggac gaaagtgttc caacatttct
     2281 attggagatg aggttcaatt tgaaattagc ataactgcta ataaatgtcc aaataaggag
     2341 tctgaaacca ttaaaattaa acctctgggc ttcactgaag aagtagaggt cgttcttcag
     2401 ttcatctgta agtgcaattg tcaaagccat ggcatcccag ccagtcccaa gtgccatgag
     2461 ggaaatggga catttgagtg tggagcctgc aggtgcaatg aggggcgtgt tgggaggcac
     2521 tgtgaatgta gcacagatga agtgaacagt gaagacatgg acgcttactg caggaaagag
     2581 aacagttcgg aaatctgcag taataatgga gaatgtgtct gtggacagtg tgtgtgtagg
     2641 aagagagata atacaaatga aatttactct ggaaaattct gcgagtgtga taacttcaac
     2701 tgtgataggt ctaatggctt aatttgtgga ggcaatggcg tgtgcaggtg tcgtgtttgt
     2761 gaatgctatc ccaattacac tggcagtgca tgtgactgtt ctttggacac tggtccatgt
     2821 ctagcgtcaa atggtcagat ctgcaatggc cggggtattt gtgaatgtgg tgcttgtaag
     2881 tgcacagatc ccaagtttca agggccaact tgtgagacat gtcagacctg ccttggcgtc
     2941 tgtgcagagc ataaagaatg tgttcagtgc agagccttca ataaaggaga aaagaaagac
     3001 acgtgtgcac aggagtgctc ccacttcaat ctcaccaaag tagaaagcag ggagaagttg
     3061 ccccagccgg tgcaggtcga tcctgtgacc cattgcaagg agaaggacat tgatgactgc
     3121 tggttctatt tcacctattc agtgaatggc aacaatgaag ctatcgtgca tgttgtggag
     3181 actccagact gtcctactgg tcccgacatc atcccaattg tagcaggcgt ggttgctgga
     3241 attgttctta ttggccttgc cttgctgctg atttggaaac ttttaatgat aattcatgac
     3301 agaagggaat ttgctaaatt tgaaaaggag aaaatgaatg ccaagtggga cacgggtgaa
     3361 aatcctattt acaagagtgc cgtgacaact gtggtcaatc cgaagtatga gggaaaagac
     3421 gtgggtggcg gagggagcga aggcggcgga tccggagggc ctggctctgg cggagagggg
     3481 agcgctggag gcgggtcagc cggtggtggc agtgtgtcaa agggagaaga gttgtttacc
     3541 ggtgttgttc ccatactggt ggaactggat ggtgacgtca atggacacaa gtttagcgtt
     3601 tctggcgaag gagaaggcga tgctacctat ggcaaactga ccctcaaact gatctgtacg
     3661 actggaaagt tgcctgtacc ttggccaaca ctggtcacaa ccttgggcta tgggctgcaa
     3721 tgctttgccc gttatcctga tcacatgaag caacatgact tctttaagtc tgctatgcca
     3781 gagggatatg tgcaggaacg gactatcttc ttcaaggacg acggcaacta caagacgaga
     3841 gctgaggtga aatttgaggg tgacacactg gtgaatcgca tagagctgaa aggcatcgac
     3901 ttcaaagaag atgggaatat tctcgggcat aaactcgagt acaactacaa ttcccacaac
     3961 gtgtacatca ctgccgataa gcagaagaat gggattaagg cgaacttcaa gatccgacac
     4021 aacatcgagg atggtggggt ccagcttgcc gaccattatc agcagaacac acccattggg
     4081 gatggaccag tcctgttgcc ggataaccac tacctcagtt accagagcaa actgtccaaa
     4141 taagaattca gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     4201 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4261 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4321 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     4381 tagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     4441 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     4501 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     4561 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     4621 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     4681 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     4741 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4801 gacggcgacg taaacggcca caagttcagc gtgaggggcg agggcgaggg cgatgccacc
     4861 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4921 accctcgtga ccaccctgag ccacggcgtg cagtgcttcg cccgctaccc cgaccacatg
     4981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5041 ttcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5101 ctggtgaacc gcatcgagct gaagggcgtc gacttcaagg aggacggcaa catcctgggg
     5161 cacaagctgg agtacaactt caacagccac aacatctata tcatggccgt caagcagaag
     5221 aacggcatca aggtgaactt caagatccgc cacaacgtgg aggacggcag cgtgcagctc
     5281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacagc
     5341 cactacctga gcacccagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg
     5401 gtcctgctgg agttccgcac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5461 taaaccggtg ctcgaggtac cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag
     5521 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     5581 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     5641 ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5701 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5761 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5821 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5881 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5941 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6001 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     6061 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6121 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6181 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     6241 ggaattcgag ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6301 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6361 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6421 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct
     6481 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6541 ttgttatccg ctaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     6601 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     6661 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     6721 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     6781 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     6841 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     6901 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     6961 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     7021 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     7081 cggccggccg ccggcgtcga cgcgcgcggc cgcggcgcgc cgttgctggc gtttttccat
     7141 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7201 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7321 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7561 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7741 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7801 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7861 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7921 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7981 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8041 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8101 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8161 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8221 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8281 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8341 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8401 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8461 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8521 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8581 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8641 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8701 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8761 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8821 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8881 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8941 aggccctttc gtctacgcgc gcgccggcgg cgcgccgtcg acggccggcc gcggccgcg
//
