LOCUS       sequence-386380-        5370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid encoding a modified version (V150A) of N-Venus, but 
            without a coiled-coil to drive reconstitution..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5370)
  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 5370)
  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
FEATURES             Location/Qualifiers
     source          1..5370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      43..70
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        89..180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             349..1008
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVILDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(415..434)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1150..1605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1237..1256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1447..1468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2292..2309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2426..2971)
                     /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(2506..2525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3066
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3246..4124)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4151..4435
                     /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     4358..4377
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4481..4498
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4562..4583)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4823..4842)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5065..5082)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5235..5321
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
       61 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
      121 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagact
      181 atgagtattc aacatttccg tgtctgctta cataaacagt aatacagggg tgtttactag
      241 aggttgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
      301 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
      361 atcacgggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
      421 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
      481 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
      541 cgcctgatga acgctcaccc ggagtttcgt atggccatga aagacggtga gctggtgatc
      601 ttggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcgtcc
      661 ctctggagtg aataccacga cgatttccgg cagtttctcc acatatattc gcaagatgtg
      721 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttt
      781 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
      841 aacttcttcg cccccgtttt cacgatgggc aaatattata cgcaaggcga caaggtgctg
      901 atgccgctgg cgatccaggt tcatcatgcc gtttgtgatg gcttccatgt cggccgcatg
      961 cttaatgaat tacaacagta ctgtgatgag tggcagggcg gggcgtaata atactagctc
     1021 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     1081 acttcgcgtt tgccacctga cgtctaagct gtcagaccaa gtttactcat atatacttta
     1141 gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1201 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1261 ttctcgccac gttcgccggc tttccccatc aagctctaaa tcgggggctc actttagggt
     1321 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     1381 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1441 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     1501 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1561 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg
     1621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgtgcgta
     1741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1861 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1921 tacctcgctc tgctaatcct gttaccagtc aggcatttga gaagcacacg gtcacactgc
     1981 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     2041 gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt
     2101 atcacttatt caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa
     2161 gctaccggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     2221 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     2281 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     2341 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     2401 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     2461 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     2521 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     2581 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     2641 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     2701 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     2761 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     2821 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     2881 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     2941 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     3001 tttctaggat catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     3061 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     3121 agaggatctg ctcatgtttg acagcttatc atcgatgcat aatgtgcctg tcaaatggac
     3181 gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt
     3241 accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
     3301 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
     3361 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
     3421 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
     3481 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
     3541 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
     3601 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
     3661 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
     3721 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
     3781 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
     3841 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
     3901 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
     3961 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
     4021 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     4081 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg
     4141 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
     4201 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
     4261 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
     4321 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
     4381 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
     4441 ttttttgggc taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg
     4501 tatggctagc gccggcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4561 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4621 gggcgatgcc acctacggca agctgaccct gaagctcatc tgcaccaccg gcaagctgcc
     4681 cgtgccctgg cccaccctcg tgaccaccct cggctacggc ctgcagtgct tcgcccgcta
     4741 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4801 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4861 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4921 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgcct atatcaccgc
     4981 cggctctggc tctggtagcg gtctagaggg tagtggctga ggtaccaagc ttggctgttt
     5041 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     5101 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     5161 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     5221 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     5281 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     5341 acgttgcgaa gcaacggccc ggagggtggc
//
