LOCUS       sequence-173936-        5216 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 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-CrN173
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..31
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      95..104
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(101..103,104..106,107..619)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(146..167)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(407..426)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    630..1252
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(829..848)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1276..1296)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1281..1610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1461..1596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1648..1667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1649..1667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1681..1697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1681..1697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1694..1716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1705..1721
                     /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(join(1729..1735,1736..1753,1754..1759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1774..1795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2083..2671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2163..2182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2842..3633,3634..3702))
                     /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     3465..3484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3703..3807)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3834..4289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3921..3940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4131..4152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4415..4437
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4429..4446
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4430..4446
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4607..4986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4987..5190
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5141..5161
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 taccatggac tacaaagacg atgacgacaa gcttgcggcc gcgaattcat cgatagatct
       61 gatatcggta ccagtcgact ctagaagatc cagcgccacc atggtgagca agggcgagga
      121 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      181 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      241 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg
      301 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      361 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      421 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      481 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat
      541 cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
      601 gatccgccac aacatcgagt agggatcccg ggtggcatcc ctgtgacccc tccccagtgc
      661 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
      721 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
      781 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
      841 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
      901 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
      961 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     1021 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     1081 gaaccactgc tcccttccct gtccttctga ttttaaaata actataccag caggaggacg
     1141 tccagacaca gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc
     1201 ttggcactgt cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc
     1261 ggccagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1381 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1441 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1501 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     1561 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg
     1621 aggaactgaa aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat
     1681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     3841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3961 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4021 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4141 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4201 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4261 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg
     4321 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4381 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4441 gccagtgcca agctgatcta tacattgaat caatattggc aattagccat attagtcatt
     4501 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     4561 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg
     4621 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     4681 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     4741 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     4801 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4861 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4921 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4981 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     5041 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     5101 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     5161 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagaa ttgatc
//
