LOCUS       sequence-386392-        5456 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes a modified version (V150A) of N-Venus followed by the 
            parallel coiled-coil CC-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 Apr 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-CC-Di
FEATURES             Location/Qualifiers
     source          1..5456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      49..135
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      227..254
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        273..364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             533..1192
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVILDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(599..618)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1334..1789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1421..1440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1631..1652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2476..2493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2610..3155)
                     /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(2690..2709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3231..3250
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3430..4308)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4335..4619
                     /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     4542..4561
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4665..4682
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4746..4767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5007..5026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5335..5352)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
       61 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      121 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
      181 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
      241 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
      301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      361 gactatgagt attcaacatt tccgtgtctg cttacataaa cagtaataca ggggtgttta
      421 ctagaggttg atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact
      481 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
      541 aaaaatcacg ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      601 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      661 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      721 tgcccgcctg atgaacgctc acccggagtt tcgtatggcc atgaaagacg gtgagctggt
      781 gatcttggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      841 gtccctctgg agtgaatacc acgacgattt ccggcagttt ctccacatat attcgcaaga
      901 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      961 ttttgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1021 ggacaacttc ttcgcccccg ttttcacgat gggcaaatat tatacgcaag gcgacaaggt
     1081 gctgatgccg ctggcgatcc aggttcatca tgccgtttgt gatggcttcc atgtcggccg
     1141 catgcttaat gaattacaac agtactgtga tgagtggcag ggcggggcgt aataatacta
     1201 gctccggcaa aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     1261 gattacttcg cgtttgccac ctgacgtcta agctgtcaga ccaagtttac tcatatatac
     1321 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1381 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1441 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcacttta
     1501 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     1561 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1621 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     1681 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1741 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     1801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2041 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2101 tacatacctc gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca
     2161 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     2221 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     2281 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
     2341 aaaagctacc ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag
     2401 tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag
     2461 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     2521 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     2581 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     2641 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     2701 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     2761 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     2821 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     2881 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc
     2941 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     3001 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     3061 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     3121 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     3181 aatatttcta ggatcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     3241 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     3301 gcgtagagga tctgctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat
     3361 ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt
     3421 cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg
     3481 gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc
     3541 gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag
     3601 cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg
     3661 gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat
     3721 atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt
     3781 atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc
     3841 aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat
     3901 ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt
     3961 attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta
     4021 aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc
     4081 tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac
     4141 caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc
     4201 gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc
     4261 attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact
     4321 cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt
     4381 cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta
     4441 acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag
     4501 aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc
     4561 cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata
     4621 cccgtttttt gggctaacag gaggaattaa ccatgggggg ttctcatcat catcatcatc
     4681 atggtatggc tagcgccggc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     4741 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4801 gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc accggcaagc
     4861 tgcccgtgcc ctggcccacc ctcgtgacca ccctcggcta cggcctgcag tgcttcgccc
     4921 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4981 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5041 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5101 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gcctatatca
     5161 ccgccggctc tggctctggt agcggtctag agggtagtgg cgaaattgct gcgctgaaac
     5221 aggaaatcgc tgccctgaaa aaagagaatg ccgctctgaa atgggaaatc gcagcgctga
     5281 aacaaggtta ggtaccaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     5341 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     5401 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccg
//
