LOCUS       sequence-386376-        5456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a modified version (V150A) of N-Venus followed by the 
            antiparallel coiled-coil apCC-Di..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5456)
  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 5456)
  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: pVRC-N-Venus-V150A-apCC-Di
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      47..74
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        93..184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             353..1012
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVILDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(419..438)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1154..1609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1241..1260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1451..1472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2296..2313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2430..2975)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2510..2529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3051..3070
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3250..4128)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4155..4439
                     /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     4362..4381
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4485..4502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4566..4587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4827..4846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5155..5172)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5325..5411
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
       61 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
      121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      181 gactatgagt attcaacatt tccgtgtctg cttacataaa cagtaataca ggggtgttta
      241 ctagaggttg atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact
      301 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
      361 aaaaatcacg ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      421 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      481 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      541 tgcccgcctg atgaacgctc acccggagtt tcgtatggcc atgaaagacg gtgagctggt
      601 gatcttggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      661 gtccctctgg agtgaatacc acgacgattt ccggcagttt ctccacatat attcgcaaga
      721 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      781 ttttgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      841 ggacaacttc ttcgcccccg ttttcacgat gggcaaatat tatacgcaag gcgacaaggt
      901 gctgatgccg ctggcgatcc aggttcatca tgccgtttgt gatggcttcc atgtcggccg
      961 catgcttaat gaattacaac agtactgtga tgagtggcag ggcggggcgt aataatacta
     1021 gctccggcaa aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     1081 gattacttcg cgtttgccac ctgacgtcta agctgtcaga ccaagtttac tcatatatac
     1141 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1201 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1261 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcacttta
     1321 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     1381 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1441 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     1501 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1561 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca
     1981 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     2041 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     2101 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
     2161 aaaagctacc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     2221 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     2281 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2341 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     2401 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     2461 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2521 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     2581 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     2641 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     2761 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     2821 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     2881 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     2941 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     3001 aatatttcta ggatcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     3061 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     3121 gcgtagagga tctgctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat
     3181 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     3241 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     3301 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     3361 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     3421 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     3481 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     3541 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     3601 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     3661 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     3721 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     3781 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     3841 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     3901 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     3961 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     4021 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     4081 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     4141 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
     4201 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
     4261 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
     4321 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
     4381 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
     4441 cccgtttttt gggctaacag gaggaattaa ccatgggggg ttctcatcat catcatcatc
     4501 atggtatggc tagcgccggc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     4561 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4621 gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc accggcaagc
     4681 tgcccgtgcc ctggcccacc ctcgtgacca ccctcggcta cggcctgcag tgcttcgccc
     4741 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4801 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4861 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4921 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gcctatatca
     4981 ccgccggctc tggctctggt agcggtctag agggtagtgg ccagctggaa caagagttgg
     5041 ctgcgctgga ccaacagatt gccgcgctga aacagcgtcg cgcagctctg aaatggcaga
     5101 tccaaggtta ggtaccaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     5161 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     5221 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     5281 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     5341 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     5401 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggaggg
//
