LOCUS       sequence-332730-        5962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-fragment of the DiB2‐split protein. This plasmid needs to be 
            co-transformed with Plasmid 168257: pMRBad-C-wtBlc to obtain 
            DiB2‐split protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5962)
  AUTHORS   Bozhanova NG, Gavrikov AS, Mishin AS, Meiler J
  TITLE     DiB-splits: nature-guided design of a novel fluorescent labeling 
            split system.
  JOURNAL   Sci Rep. 2020 Jul 6;10(1):11049. doi: 10.1038/s41598-020-67095-2.
  PUBMED    32632329
REFERENCE   2  (bases 1 to 5962)
  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: "Sci Rep."; 
            date: "2020-07-6"; volume: "10(1):11049. doi"; pages: " 
            10.1038/s41598-020-67095-2"
COMMENT     Sequence Label: pET11a-N-DiB2
FEATURES             Location/Qualifiers
     source          1..5962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..26
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(377..395)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      391..438
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(613..618,619..635,636..641))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(668..686)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        754..858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(859..927,928..1719)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1077..1096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1890..2478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2379..2398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2632..2649
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2664..2803
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2729..2748)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2889..2911
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2905..3096)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4408..5490)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5452..5471
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5491..5568)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5774..5793
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5877..5896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5877..5895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5896..5920
                     /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)."
     RBS             5935..5957
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atatggggca tcaccaccac catcacctcg aatcaaccag cttgtacaag aaatccagct
       61 ccacgccgcc gcgtggcgtg accgtagtaa ataatttcga ctgcaaacgt tatctcggta
      121 cctggtatga gattgcccgt tttgatcacc gctttgaacg tggactggaa aaagtcaccg
      181 caacatacag cctgcgtgat gacggcggcc tgaatgtcat taataaaggc tataaccctg
      241 acagaggaat gtggcagcag agtgaaggga aagcgtactt taccggcgca ccaactcgcg
      301 ctgcgctgaa agtgtcattc ttttaaggat ccggctgcta acaaagcccg aaaggaagct
      361 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      421 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg
      481 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
      541 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
      601 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
      661 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
      721 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      841 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1081 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1381 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     1441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1561 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2521 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2581 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2641 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2701 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2761 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     2821 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2881 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2941 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3001 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3061 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3121 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3181 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3241 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3301 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     3361 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     3421 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     3481 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     3541 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     3601 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     3661 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     3721 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     3781 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     3841 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
     3901 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     3961 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     4021 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     4081 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4141 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     4201 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4261 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4321 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4381 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4441 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4501 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     4561 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4621 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4681 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4741 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4801 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4861 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4921 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4981 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5041 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5101 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5161 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5221 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5281 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5341 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5401 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     5461 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     5521 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     5581 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     5641 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5701 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5761 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5821 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     5881 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5941 taactttaag aaggagatat ac
//
