LOCUS       Exported                7488 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  DULIP negative control (bait) to exclude unspecific background 
            binding.
ACCESSION   .
VERSION     .
KEYWORDS    PA-RL-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7488)
  AUTHORS   Trepte P, Buntru A, Klockmeier K, Willmore L, Arumughan A, Secker C,
            Zenkner M, Brusendorf L, Rau K, Redel A, Wanker EE
  TITLE     DULIP: A Dual Luminescence-Based Co-Immunoprecipitation Assay for 
            Interactome Mapping in Mammalian Cells.
  JOURNAL   J Mol Biol. 2015 Oct 23;427(21):3375-88. doi: 
            10.1016/j.jmb.2015.08.003. Epub 2015 Aug 8.
  PUBMED    26264872
REFERENCE   2  (bases 1 to 7488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7488
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      940..949
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             970..1143
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1147..1317
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     CDS             1333..2265
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    2278..2302
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2305..3012
                     /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(2450..2468)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2718..2737
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3014..3038)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3084..3101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3090..3314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3360..3788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3447..3466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3657..3678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3797..3817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3802..4131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3982..4117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4044..4063
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4198..4992
                     /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(4252..4271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4862..4881
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5166..5287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5203..5222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5257..5276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5336..5352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5336..5352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5349..5371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5360..5376
                     /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(5384..5414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6494..7354)
                     /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     7117..7136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7355..7459)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgg taccgagctc ggatccgccg ccaccatggt tgcgcaacac
      961 gatgaagccg tggacaacaa attcaacaaa gaacaacaaa acgcgttcta tgagatctta
     1021 catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt aaaagatgac
     1081 ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgatgc tcaggcgccg
     1141 aaagtagaca acaaattcaa caaagaacaa caaaacgcgt tctatgagat cttacattta
     1201 cctaacttaa acgaagaaca acgaaacgcc ttcatccaaa gtttaaaaga tgacccaagc
     1261 caaagcgcta accttttagc agaagctaaa aagctaaatg gtgctcaggc gccgaaagta
     1321 gacgcgaatt cgatggcttc caaggtgtac gaccccgagc aacgcaaacg catgatcact
     1381 gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt catcaactac
     1441 tatgattccg agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc
     1501 agctacctgt ggaggcacgt cgtgcctcac atcgagcccg tggctagatg catcatccct
     1561 gatctgatcg gaatgggtaa gtccggcaag agcgggaatg gctcatatcg cctcctggat
     1621 cactacaagt acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt
     1681 gtgggccacg actggggggc ttgtctggcc tttcactact cctacgagca ccaagacaag
     1741 atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg
     1801 cctgacatcg aggaggatat cgccctgatc aagagcgaag agggcgagaa aatggtgctt
     1861 gagaataact tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa actggagcct
     1921 gaggagttcg ctgcctacct ggagccattc aaggagaagg gcgaggttag acggcctacc
     1981 ctctcctggc ctcgcgagat ccctctcgtt aagggaggca agcccgacgt cgtccagatt
     2041 gtccgcaact acaacgccta ccttcgggcc agcgacgatc tgcctaagat gttcatcgag
     2101 tccgaccctg ggttcttttc caacgctatt gtcgagggag ctaagaagtt ccctaacacc
     2161 gagttcgtga aggtgaaggg cctccacttc agccaggagg acgctccaga tgaaatgggt
     2221 aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg agcagctcga ggatatcaca
     2281 agtttgtaca aaaaagcagg cttcatggtg agcaagggcg aggaggataa catggccatc
     2341 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     2401 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2461 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     2521 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     2581 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     2641 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     2701 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     2761 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2821 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2881 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2941 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     3001 gagctgtaca aggacccagc tttcttgtac aaagtggtga tatcgcggcc gctctagagg
     3061 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3121 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3181 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3241 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3301 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3541 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3661 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3721 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3781 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3961 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4021 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4081 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4141 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4861 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4921 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4981 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5041 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5101 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5161 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5221 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5281 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6301 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6361 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6421 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6481 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6541 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6601 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6661 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6721 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6781 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6841 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6901 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6961 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7021 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7081 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7141 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7201 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7261 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7321 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7381 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7441 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacg
//
