LOCUS       sequence-457434-        4215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  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
REFERENCE   2  (bases 1 to 4215)
  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"; journalName: "Nat Protoc.
            2008  . 3(4):588-96."
COMMENT     Sequence Label: pCE-BiFC-VC155
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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     227..249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     246..262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     246..262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             827..853
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             935..1186
                     /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     1123..1144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2046..2068)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2168..2187
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2265..2283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2351..2455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2456..2524,2525..3316)
                     /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(2674..2693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3487..4075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3976..3995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
       61 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc
      301 tagaggatca ccaaaaacgg aacgttgagc tggacggaaa tagtggtaaa gtgacatgat
      361 tatagtttga agatttctaa tttcacaatt agagcaaatg ttgttcggta tttattttca
      421 acggtattta tactattttc cacctttttc tagaacattc gagctgcttg ttgcaaaagg
      481 agggcgactc acattcggta catggaaaag tagtgtacac aataaagaga cccagataca
      541 ttttccgtct gcgtctcttt gcacccaccg ggagtatttt caaacgaatg catctaggac
      601 cttctagaac attctgtaag gctgcagaat gcgggtatat aaggaaagcg ggctcagagg
      661 aagccaacac gctttgttct agtgcatcta aaaaacttcg aaaatcctca tcggatcccc
      721 gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt
      781 ttcaggagga cccttggcta gcaattgtac ccgcgggccc accatgtacc catacgatgt
      841 tccagattac gctcttatgg ccatggaggc ccgaattcgg tcgaccgaga tctctcgagg
      901 taccggaggt ggcgggagcg gaggtggcgg gagtgacaag cagaagaacg gcatcaaggc
      961 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1021 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1081 ccagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1141 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgcagg
     1201 atccagacat gataagatac gatatctgag ctccgcatcg gccgctgtca tcagatcgcc
     1261 atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc ctacatgctc
     1321 tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct cttaatttct
     1381 ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag
     1441 aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt aataataatt
     1501 ctatcccaaa atctacacaa tgttctgtgt acacttctta tgttttttac ttctgataaa
     1561 tttttttgaa acatcataga aaaaaccgca cacaaaatac cttatcatat gttacgtttc
     1621 agtttatgac cgcaattttt atttcttcgc acgtctgggc ctctcatgac gtcaaatcat
     1681 gctcatcgtg aaaaagtttt ggagtatttt tggaattttt caatcaagtg aaagtttatg
     1741 aaattaattt tcctgctttt gctttttggg gtttccccta ttgtttgtca agatttcgag
     1801 gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag aaagatcgga
     1861 agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt tgaaagtgga
     1921 gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat accaaacata
     1981 actgtttcta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2041 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2101 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2161 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2221 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     2281 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2341 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2401 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2461 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2521 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2581 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2641 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2701 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2761 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2821 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2881 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2941 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3001 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3061 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3121 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3181 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3241 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3301 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3601 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3781 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4081 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4141 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4201 cgctcgccgc agccg
//
