LOCUS       sequence-438019-        2731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiGreen 1.0 HG entry vector containing filler sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2731)
  AUTHORS   Pennetti VJ, LaFayette PR, Parrott WA
  TITLE     MultiGreen: A multiplexing architecture for GreenGate cloning.
  JOURNAL   PLoS One. 2024 Sep 18;19(9):e0306008. doi: 
            10.1371/journal.pone.0306008. eCollection 2024.
  PUBMED    39292669
REFERENCE   2  (bases 1 to 2731)
  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: "PLoS One.";
            date: "2024-09-18"; volume: "19(9):e0306008. doi"; pages: " 
            10.1371/journal.pone.0306008. eCollection 2024"
COMMENT     Sequence Label: pVP088
FEATURES             Location/Qualifiers
     source          1..2731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..96)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(78..96)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(106..128)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(322..341)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     441..463
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(501..519)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        587..691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             876..1254
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(910..929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1716..2304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2205..2224
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2458..2475
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2592..2613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2628..2633,2634..2651,2652..2658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2666..2682
                     /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        2696..2714
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2696..2713
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taggacctag agacgagtcg aggacgtgcc agctcaacga gtggccgtct ctgtatgtat
       61 tgagaccgaa ttctcgccct atagtgagtc gtattacctg gccgtcgttt tacaacgtcg
      121 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      181 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      241 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      301 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
      361 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      421 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      481 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
      541 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      661 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
      721 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      781 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
      841 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
      901 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
      961 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1021 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1081 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1141 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1201 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     1261 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     1321 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     1381 tgataaatct ggagccggtg agcgtggggt atcattgcag cactggggcc agatggtaag
     1441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1561 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     2641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac gagctattta
     2701 ggtgacacta tagaagtgaa gcttggtctc a
//
