LOCUS       sequence-438015-        3518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRFP1e chromoprotein in a MultiGreen 1.0 HG entry vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3518)
  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 3518)
  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: pVP086
FEATURES             Location/Qualifiers
     source          1..3518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    16..37
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        38..72
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             81..92
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             99..776
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     misc_feature    805..825
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(857..876)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(858..876)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(886..908)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1102..1121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1221..1243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1281..1299)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1367..1471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1472..1540,1541..2332)
                     /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(1690..1709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2503..3091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2992..3011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3245..3262
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3379..3400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3415..3420,3421..3438,3439..3445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3453..3469
                     /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        3483..3501
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3483..3500
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taggacctag agacggaatt cgcggccgct tctagagttt acggctagct cagtcctagg
       61 tacaatgcta gctactagag aaagaggaga aatactagat ggcttcctcc gaagacgtta
      121 tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg
      181 aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag
      241 ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg
      301 gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc
      361 cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta
      421 cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca
      481 acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg
      541 aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga
      601 aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc
      661 agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact
      721 acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac
      781 gctgatagtg ctagtgtaga tcgctactag tagcggccgc tgcagcgtct ctgtatgtat
      841 tgagaccgaa ttctcgccct atagtgagtc gtattacctg gccgtcgttt tacaacgtcg
      901 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      961 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     1021 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     1081 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     1141 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     1201 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     1261 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     1321 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1381 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1441 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1501 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1561 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1621 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1681 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1741 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1801 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1861 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     1921 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     1981 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2041 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2101 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2161 tgataaatct ggagccggtg agcgtggctc tcgaggtatc attgcagcac tggggccaga
     2221 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2281 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2341 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2401 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2461 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2521 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2581 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2641 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2701 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2761 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2821 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2881 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2941 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3001 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3061 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3121 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3181 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3241 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3301 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3361 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     3421 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacgag
     3481 ctatttaggt gacactatag aagtgaagct tggtctca
//
