LOCUS       sequence-332732-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-fragment of the DiB-RM‐split‐Zip protein. This plasmid needs to be
            co-transformed with Plasmid 168477: pMRBad-Z-C-DiB-RM to obtain 
            DiB-RM‐split-Zip protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Bozhanova NG, Harp JM, Bender BJ, Gavrikov AS, Gorbachev DA, Baranov
            MS, Mercado CB, Zhang X, Lukyanov KA, Mishin AS, Meiler J
  TITLE     Computational redesign of a fluorogen activating protein with 
            Rosetta.
  JOURNAL   PLoS Comput Biol. 2021 Nov 8;17(11):e1009555. doi: 
            10.1371/journal.pcbi.1009555. eCollection 2021 Nov.
  PUBMED    34748541
REFERENCE   2  (bases 1 to 6073)
  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: "PLoS Comput
            Biol."; date: "2021-11-8"; volume: "17(11):e1009555. doi"; pages: " 
            10.1371/journal.pcbi.1009555. eCollection 2021 Nov"
COMMENT     Sequence Label: pET11a-Z-N-DiB-RM
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1793..1984
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1978..2000)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2086..2225
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2141..2160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2240..2257)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2411..2999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2491..2510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3170..3961,3962..4030))
                     /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     3793..3812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4031..4135)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4203..4221
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4248..4253,4254..4270,4271..4276)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(4451..4498)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4494..4512
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4590..4601)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(4974..4991)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5005..5027
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5042..5066
                     /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     complement(5066..5085)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5067..5085)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5169..5188)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5394..5471
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5472..6073,1..481)
                     /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     complement(5491..5510)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
       61 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
      121 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
      181 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
      241 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
      301 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
      361 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
      541 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
      601 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
      661 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
      721 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
      781 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
      841 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
      901 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
      961 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     1021 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     1081 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     1141 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     1201 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     1261 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     1321 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     1381 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     1441 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     1501 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     1561 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     1621 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     1681 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     1741 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     1801 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     1861 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     1921 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     1981 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2041 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2101 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     2161 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2221 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2281 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2341 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2401 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2461 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2521 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2581 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2641 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2701 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2761 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2821 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2881 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2941 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3001 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3061 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3241 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3301 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3361 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3421 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3481 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3541 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3601 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3661 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3721 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3781 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3841 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3901 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3961 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4021 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4081 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4141 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4201 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt ctcatgtttg acagcttatc
     4261 atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     4321 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
     4381 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatccg gatatagttc
     4441 ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt
     4501 attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat
     4561 ccttactgcg ccagttcctt tttcagagct tgtaactccc acttcagctg cgcaagttct
     4621 tttttgtttg cctgcaattc ttttttgagg gcgctcgagc cagagccaga gccaccaaag
     4681 aacgccactt tcagcgcagc gcgagttggt gcgccggtaa agtacgcttt cccaacgtac
     4741 tgctgccaca ttcctctgtc agggttatag cctttgaaga tgacattcag gccgccgtca
     4801 tcacgcaggc tgtatgttgc ggtgactttt tccagtccac gttctgcgcg gtgatcaaaa
     4861 cgggcaatct cataccaggt accgagataa cgtttgcagt cgaaattatt tactacggtc
     4921 acgccacgcg gcggcgtgga gctggatttc ttgtacaagc tggttgattc gaggtgatgg
     4981 tggtggtgat gacccatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
     5041 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
     5101 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     5161 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     5221 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     5281 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     5341 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     5401 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     5461 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     5521 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     5581 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     5641 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     5701 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     5761 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     5821 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     5881 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     5941 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     6001 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     6061 gtctggctgg ctg
//
