LOCUS       sequence-437922-        2716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiGreen 1.0 FH entry vector containing filler sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2716)
  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 2716)
  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: pVP090
FEATURES             Location/Qualifiers
     source          1..2716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(56..74)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(84..106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(300..319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     419..441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(479..497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        565..669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(670..738,739..1530)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(888..907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1701..2289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2190..2209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2443..2460
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2577..2598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2613..2618,2619..2636,2637..2643)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2651..2667
                     /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        2681..2699
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2681..2698
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 actaagttgg catgaagcta ctatatagaa gcattaggtg agaccgaatt ctcgccctat
       61 agtgagtcgt attacctggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      301 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      421 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
      481 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
      541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1381 cgtggctctc gaggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2341 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2401 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2461 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2521 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2581 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     2641 ttgtgtggaa ttgtgagcgg ataacaattt cacacgagct atttaggtga cactatagaa
     2701 gtgaagcttg gtctca
//
