LOCUS       sequence-354487-        5216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a fluorescent protein fragment derived from 
            Venus for BiFC and multicolor BiFC assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5216)
  AUTHORS   Shyu YJ, Liu H, Deng X, Hu CD
  TITLE     Identification of new fluorescent protein fragments for bimolecular 
            fluorescence complementation analysis under physiological 
            conditions.
  JOURNAL   Biotechniques. 2006 Jan . 40(1):61-6.
  PUBMED    16454041
REFERENCE   2  (bases 1 to 5216)
  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: 
            "Biotechniques. 2006 Jan . 40(1):61-6."
COMMENT     Sequence Label: pBiFC-VN173
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     219..239
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    263..885
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     667..686
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(join(896..1408,1409..1411,1412..1414))
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     1089..1108
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1348..1369
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1411..1420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1484..1507)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1541..1744)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1570..1590)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1745..2124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2285..2302)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2285..2301)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2294..2316)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2442..2897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2579..2600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2791..2810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2924..3028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3029..3097,3098..3889)
                     /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(3247..3266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4060..4648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4549..4568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4802..4819
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4936..4957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4972..4977,4978..4995,4996..5002)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5010..5026
                     /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     5015..5037
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5034..5050
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5034..5050
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5064..5083
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5064..5082
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(5121..5216,1..234))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5135..5216,1..54)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
       61 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      121 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
      181 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
      241 cagccaagct ggccgcgtac ccaattcaac aggcatctac tgagtggacc caacgcatga
      301 gaggacagtg ccaagcaagc aactcaaatg tcccaccggt tgggcatggc caggtagcct
      361 atgctgtgtc tggacgtcct cctgctggta tagttatttt aaaatcagaa ggacagggaa
      421 gggagcagtg gttcacgcct gtaatcccag caatttggga ggccaaggtg ggtagatcac
      481 ctgagattag gagttggaga ccagcctggc caatatggtg aaaccccgtc tctaccaaaa
      541 aaacaaaaat tagctgagcc tggtcatgca tgcctggaat cccaacaact cgggaggctg
      601 aggcaggaga atcgcttgaa cccaggaggc ggagattgca gtgagccaag attgtgccac
      661 tgcactccag cttggttccc aatagacccc gcaggcccta caggttgtct tcccaacttg
      721 ccccttgctc cataccaccc ccctccaccc cataatatta tagaaggaca cctagtcaga
      781 caaaatgatg caacttaatt ttattaggac aaggctggtg ggcactggag tggcaacttc
      841 cagggccagg agaggcactg gggaggggtc acagggatgc cacccgggat ccctactcga
      901 tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat
      961 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     1021 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     1081 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     1141 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
     1201 agcactgcag gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     1261 cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     1321 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     1381 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gatggatctt ctagagtcga
     1441 ctggtaccga tatcagatct atcgatgaat tcgcggccgc aagcttgtcg tcatcgtctt
     1501 tgtagtccat ggtagatcaa ttctgacggt tcactaaacg agctctgctt atatagacct
     1561 cccaccgtac acgcctaccg cccatttgcg tcaacggggc ggggttatta cgacattttg
     1621 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     1681 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt ggtgtactgc caaaaccgca
     1741 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccgta
     1801 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     1861 gcggacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     1921 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     1981 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     2041 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     2101 aataactagt caataatcaa tgtcaacatg gcggtcatat tggacatgag ccaatataaa
     2161 tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata ttgatttatg
     2221 ctatataacc aatgactaat atggctaatt gccaatattg attcaatgta tagatcagct
     2281 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2341 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2401 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat
     2461 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2521 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2881 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2941 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3001 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3061 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3121 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3181 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3241 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3301 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3361 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3421 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3481 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3541 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3601 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3661 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3721 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3781 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3841 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3901 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3961 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4201 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4261 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4321 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4381 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4441 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4501 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4561 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4621 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4681 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4741 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4801 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4861 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4921 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4981 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5041 cagctatgac atgattacga atttaatacg actcactata gggaattaac tttctggttt
     5101 ttcagttcct cgaggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     5161 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaat
//
