LOCUS       sequence-321691-        7712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein interaction screening tool with Gateway-compatible BiFC 
            vectors comprising of a set of 4 vectors permiting N or C fusions of
            candidate genes to the N or C fragment of Split-Venus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7712)
  AUTHORS   Lepur A, Kovacevic L, Beluzic R, Vugrek O
  TITLE     Combining Unique Multiplex Gateway Cloning and Bimolecular 
            Fluorescence Complementation (BiFC) for High-Throughput Screening of
            Protein-Protein Interactions.
  JOURNAL   J Biomol Screen. 2016 Dec;21(10):1100-1111. doi: 
            10.1177/1087057116659438. Epub 2016 Jul 28.
  PUBMED    27455993
REFERENCE   2  (bases 1 to 7712)
  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"; doi: 
            "10.1177/1087057116659438"; journalName: "J Biomol Screen"; date: 
            "2016-12"; volume: "21"; issue: "10"; pages: "1100-1111"
COMMENT     Sequence Label: pVenusCfC
FEATURES             Location/Qualifiers
     source          1..7712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(834..938)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1048..1351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1352..1555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1506..1526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1552..1576
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1673..1797
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1822..1827,1828..1845,1846..1852)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1906..2565
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1972..1991)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2907..3212
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3091..3110
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3253..3377)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     3582..3603
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3674..3693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3675..3693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3704..3721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3704..3720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3713..3735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    4125..4259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4162..4181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4216..4235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4385..4840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4472..4491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4682..4703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4957..5314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4957..5148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4980..5000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5165..5300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5365..6159
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDPLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5419..5438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6029..6048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6223..6271
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6312..6331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6431..6453
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6577..6681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6682..6750,6751..7542)
                     /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     complement(6900..6919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagagctt gcaattcgcg
      841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      961 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtattacg
     1021 aggccctttc acgcattaga tgcatgtcgt tacataactt acggtaaatg gcccgcctgg
     1081 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     1141 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1201 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     1261 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     1321 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     1381 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1441 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     1501 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     1561 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
     1621 ccgggaccga tccagcctcc ggactctagc ctaggccgcg gagctcgagt caacaagttt
     1681 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     1741 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     1801 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     1861 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1921 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1981 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     2041 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     2101 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     2161 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     2221 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     2281 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     2341 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2401 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2461 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2521 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc
     2581 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     2641 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     2701 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     2761 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     2821 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     2881 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     2941 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     3001 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     3061 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     3121 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     3181 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     3241 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     3301 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     3361 ttcttgtaca aagtggttga taccggtcgg agtatagcca ccaagaacgg catcaaggcc
     3421 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     3481 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     3541 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3601 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagacgcg tgcatgagac
     3661 gtcatagctc tctccctata gtgagtcgta ttataagcta ggcactggcc gtcgttttac
     3721 aacgtcgtga ctgggaaaac tgctagcttg ggatctttgt gaaggaacct tacttctgtg
     3781 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
     3841 ttttaagtgt ataatgtgtt aaactagctg catatgcttg ctgcttgaga gttttgctta
     3901 ctgagtatga tttatgaaaa tattatacac aggagctagt gattctaatt gtttgtgtat
     3961 tttagattca cagtcccaag gctcatttca ggcccctcag tcctcacagt ctgttcatga
     4021 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4081 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4141 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4201 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     4261 atcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggctggcgt
     4321 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4381 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     4861 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     4921 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     4981 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5041 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     5101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5161 gcccatcccg cccctaactc cgcccaattc cgcccattct ccgccccatg gctgactaat
     5221 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5281 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     5341 ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg
     5401 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5461 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5521 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5581 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggattggctg
     5641 ctattgggcg aagtgccggg gcaggatccc ctgtcatctc accttgctcc tgccgagaaa
     5701 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5761 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5821 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5881 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5941 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6001 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6061 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6121 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6181 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     6241 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg
     6301 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6361 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6421 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6481 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6541 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6661 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6721 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6781 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6841 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6901 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6961 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7021 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7081 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7141 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7201 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7261 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7321 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7381 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7441 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7501 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7561 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7621 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7681 tcagaccccg tagaaaagat caaaggatct tc
//
