LOCUS       sequence-152662-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accessory plasmid (encodes M13 gene VI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5300)
  AUTHORS   Brodel AK, Jaramillo A, Isalan M
  TITLE     Engineering orthogonal dual transcription factors for multi-input 
            synthetic promoters.
  JOURNAL   Nat Commun. 2016 Dec 16;7:13858. doi: 10.1038/ncomms13858.
  PUBMED    27982027
REFERENCE   2  (bases 1 to 5300)
  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 
            Commun."; date: "2016-12-16"; volume: "7:13858. doi"; pages: " 
            10.1038/ncomms13858"
COMMENT     Sequence Label: pJPC12-Delta-M13-PRM-B0034-geneVI
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     471..493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     485..502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     486..502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     512..531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        512..530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     555..571
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     556..572
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             684..695
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             702..1040
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     primer_bind     complement(1092..1112)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1092..1110)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1131..1147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1131..1147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1155..1171
                     /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)."
     promoter        complement(join(1179..1185,1186..1203,1204..1209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1362..1379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      2524..2746
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2794..3744
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
       61 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
      121 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
      181 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttaca
      241 aaaggatcta ggtgaagatc ctttttgata atctcatgag cggatacata tttgaatgta
      301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccaccttcag
      361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      481 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
      541 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tcgcaaccat
      601 tatcaccgcc agaggtaaaa tagtcaacac gcacggtgtt agatattaca aactttcttg
      661 tatagattta caatgtatct tgtaaagagg agaaatacta gatgccagtt cttttgggta
      721 ttccgttatt attgcgtttc ctcggtttcc ttctggtaac tttgttcggc tatctgctta
      781 cttttcttaa aaagggcttc ggtaagatag ctattgctat ttcattgttt cttgctctta
      841 ttattgggct taactcaatt cttgtgggtt atctctctga tattagcgct caattaccct
      901 ctgactttgt tcagggtgtt cagttaattc tcccgtctaa tgcgcttccc tgtttttatg
      961 ttattctctc tgtaaaggct gctattttca tttttgacgt taaacaaaaa atcgtttctt
     1021 atttggattg ggataaataa cgcaaaaaac cccgcttcgg cggggttttt tcgcagctcc
     1081 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     1141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1441 tccacagaat caggggataa cgcaggaaag aacatgtgag atctctacgg gtcggatttg
     1501 aagtcgtctt ggtaggaggc agcctgaatg gcgaatgccg atgcccttga gagccttcaa
     1561 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     1621 cttctttatc atgcaactcg taggacaggg tgccggcagc gctctgggtc attttcggcg
     1681 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     1741 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     1801 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     1861 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     1921 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     1981 gatcgctcgc ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga
     2041 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     2101 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     2161 tggaagccgg cggcacctcg ctaacggatt caccactccg cagacccgcc ataaaacgcc
     2221 ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata
     2281 aaaggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact
     2341 gaatgtcacg aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc
     2401 agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt
     2461 tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt
     2521 agtgagttat acacagggct gggatctatt ctttttatct ttttttattc tttctttatt
     2581 ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt
     2641 acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata
     2701 cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat
     2761 agtgattaaa atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca
     2821 aatgaactag cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc
     2881 tgtgtggcac tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg
     2941 ttcacttata accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta
     3001 ttagctaaag caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt
     3061 aaaggcttta agatcttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa
     3121 ttagttttta gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat
     3181 aatctggaac atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta
     3241 ttaaaagaac taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa
     3301 tttaagttca tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg
     3361 gttttgaaac caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat
     3421 aagcgaggcc gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat
     3481 ctcgtaaccg aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc
     3541 attacatcag attcctacct acataacgga ctaagaaaaa cactacacga tgctttaaca
     3601 gcaaaaattc agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat
     3661 gatctcaatg gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata
     3721 ctggctaaat acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc
     3781 aagactaaca aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc
     3841 catatgcaca gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt
     3901 tggtgcattc aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt
     3961 aacacctaat aagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac
     4021 acctgagaca acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc
     4081 tgcttatcga atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa
     4141 aatcatggca attctggaag aaatagcgct ttcagccggc aaaccggctg aagccggatc
     4201 tgcgattctg ataacaaact agcaacacca gaacagcccg tttgcgggca gcaaaacccg
     4261 tacttttgga cgttccggcg gttttttgtg gcgagtggtg ttcgggcggt gcgcgcaaga
     4321 tccattatgt taaacgggcg agtttacatc tcaaaaccgc ccgcttaaca ccatcagaaa
     4381 tcctcagcgc gattttaagc accaaccccc ccccgtaaca cccaaatcca tactgaaagt
     4441 ggctttgttg aataaatcag atttcgggta agtctccccc gtagcgggtt gtgttttcag
     4501 gcaatacgca cgctttcagg catacctgct ttcgtcattt tgttcagcgc tcgtaccagg
     4561 gccatagcct ccgcaacctg accatcgtag tcacgcagcg tcagtgaacc cccgaacaga
     4621 gatctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agacaataac
     4681 cctgataaat gcttcaataa tcgcagacca aaacgatctc aagaagatca tcttattaat
     4741 cagataaaat atttctaggc accaataact gccttaaaaa aattacgccc cgccctgcca
     4801 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     4861 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     4921 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     4981 ggtgaaactc acccagggat tggctgaaac gaaaaacata ttctcaataa acccttttag
     5041 ggaaataagg ccaggttttc accgtaacac gccacatctt gggaaatatt tgtgtagaaa
     5101 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     5161 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     5221 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     5281 aaacttgtgc ttatttttcc
//
