LOCUS       sequence-386378-        4424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the parallel coiled-coil CC-Di followed by C-Venus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4424)
  AUTHORS   Rhys GG, Cross JA, Dawson WM, Thompson HF, Shanmugaratnam S, Savery 
            NJ, Dodding MP, Hocker B, Woolfson DN
  TITLE     De novo designed peptides for cellular delivery and subcellular 
            localisation.
  JOURNAL   Nat Chem Biol. 2022 Jul 14. pii: 10.1038/s41589-022-01076-6. doi: 
            10.1038/s41589-022-01076-6.
  PUBMED    35836017
REFERENCE   2  (bases 1 to 4424)
  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 Chem 
            Biol. 2022 Jul 14. pii: 10.1038/s41589-022-01076-6. doi: 
            10.1038/s41589-022-01076-6."
COMMENT     Sequence Label: pBAD-CC-Di-C-Venus
FEATURES             Location/Qualifiers
     source          1..4424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        542..826
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     749..768
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             872..889
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1031..1282
                     /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     1219..1240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1366..1383)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1536..1622
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1714..1741
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1760..1851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1852..1920,1921..2712)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2070..2089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2883..3471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3372..3391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3625..3642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3657..3797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3723..3742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3883..3905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(4061..4424,1..515))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
       61 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
      121 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
      181 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
      241 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
      301 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
      361 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
      421 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
      481 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
      541 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
      601 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
      661 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
      721 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
      781 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
      841 ctaacaggag gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag
      901 cctgcagggc gaaattgctg cgctgaaaca ggaaatcgct gccctgaaaa aagagaatgc
      961 cgctctgaaa tgggaaatcg cagcgctgaa acaaggtagc ggtagcggca gcggtgaggg
     1021 cctgcagggg gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1081 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1141 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1201 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1261 cggcatggac gagctgtaca agggcagcgg cagcggcagc ggtctagagg gtagtggctg
     1321 aggtaccaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1381 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1441 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1501 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1561 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1621 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1681 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1741 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3001 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3541 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3601 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3661 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     3721 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3781 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3901 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
     3961 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     4021 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     4081 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     4141 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     4201 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     4261 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     4321 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4381 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tgga
//
