LOCUS       pCE-BiFC-VC155                4224 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCE-BiFC-VC155
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  AUTHORS   Shyu YJ, Hiatt SM, Duren HM, Ellis RE, Kerppola TK, Hu CD
  TITLE     Visualization of protein interactions in living Caenorhabditis 
            elegans using bimolecular fluorescence complementation analysis.
  JOURNAL   Nat Protoc. 2008  . 3(4):588-96.
  PUBMED    18388940
COMMENT     This file is created by Vector NTI
            //gt12365.cn/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|gt12365.cn|
COMMENT     VNTNAME|pCE-BiFC-VC155|
FEATURES             Location/Qualifiers
     source          1..4224
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             112..363
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     300..321
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1231..1253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1353..1372
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1450..1468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1536..1640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1641..2501
                     /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(1859..1878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2672..3260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3161..3180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3414..3431
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3548..3569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3584..3614
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3622..3638
                     /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)."
     primer_bind     3627..3649
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3646..3662
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3646..3662
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgtacccat acgatgttcc agattacgct cttatggcca tggaggcccg aattcggtcg
       61 accgagatct ctcgaggtac cggaggtggc gggagcggag gtggcgggag tgacaagcag
      121 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      181 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      241 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
      301 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      361 aagtaagcgg ccgcaggatc cagacatgat aagatacgat atctgagctc cgcatcggcc
      421 gctgtcatca gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
      481 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
      541 aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
      601 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
      661 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
      721 gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg cacacaaaat
      781 accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg
      841 gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt
      901 ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg ggggtttccc
      961 ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     1021 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     1081 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     1141 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     1201 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1261 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1321 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1381 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     1441 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1501 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1561 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1621 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1681 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1741 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1801 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1861 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1921 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1981 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2041 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2101 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2161 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2221 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2281 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2341 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2401 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2461 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2521 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2581 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2641 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2701 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2761 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2821 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2881 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2941 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3001 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3061 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3301 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3361 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3421 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3481 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3541 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     3601 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     3661 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatca ccaaaaacgg
     3721 aacgttgagc tggacggaaa tagtggtaaa gtgacatgat tatagtttga agatttctaa
     3781 tttcacaatt agagcaaatg ttgttcggta tttattttca acggtattta tactattttc
     3841 cacctttttc tagaacattc gagctgcttg ttgcaaaagg agggcgactc acattcggta
     3901 catggaaaag tagtgtacac aataaagaga cccagataca ttttccgtct gcgtctcttt
     3961 gcacccaccg ggagtatttt caaacgaatg catctaggac cttctagaac attctgtaag
     4021 gctgcagaat gcgggtatat aaggaaagcg ggctcagagg aagccaacac gctttgttct
     4081 agtgcatcta aaaaacttcg aaaatcctca tcgggatccc cgggattggc caaaggaccc
     4141 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
     4201 agcaattgta cccgcgggcc cacc
//