LOCUS       sequence-396441-        5106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RKIP (PEBP1) expression plasmid with a RFP reporter on the backbone.
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5106)
  AUTHORS   Wan Y, Cohen J, Szenk M, Farquhar KS, Coraci D, Krzyszton R, Azukas 
            J, Van Nest N, Smashnov A, Chern YJ, De Martino D, Nguyen LC, Bien 
            H, Bravo-Cordero JJ, Chan CH, Rosner MR, Balazsi G
  TITLE     Nonmonotone invasion landscape by noise-aware control of metastasis 
            activator levels.
  JOURNAL   Nat Chem Biol. 2023 Jul;19(7):887-899. doi: 
            10.1038/s41589-023-01344-z. Epub 2023 May 25.
  PUBMED    37231268
REFERENCE   2  (bases 1 to 5106)
  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"; doi: 
            "10.1038/s41589-023-01344-z"; journalName: "Nat Chem Biol"; date: 
            "2023-07"; volume: "19"; issue: "7"; pages: "887-899"
COMMENT     Sequence Label: pCMV-RKIP_RFP
FEATURES             Location/Qualifiers
     source          1..5106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(41..59)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(42..59)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(79..96)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    85..309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   337..371
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        472..971
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(507..530)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     904..923
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1011..1020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1736..1960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2121..2709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2201..2220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2880..3671,3672..3740))
                     /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     3503..3522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3741..3845)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3872..4251
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4252..4455
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4406..4426
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4500..4519
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4500..4518
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agcagctgtc tgggaagtaa gcgactctag agggccctat tctatagtgt cacctaaatg
       61 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
      121 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
      181 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
      241 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
      301 gctctatggc ttgtcgacga aggctctctc gagatgcatg cgtcaatttt acgcagacta
      361 tctttctagg gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca
      421 gtcatcgccc aagctggcgc tatctgggca tcggggagga agaagcccgt ggggtagggg
      481 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
      541 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
      601 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
      661 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
      721 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
      781 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgtgtccg
      841 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
      901 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
      961 cgggcctttc gacctgcagc gacccgctta acagcgtcaa cagcgtgccg gccaccatgg
     1021 cccttagtaa gcacggcctg actaaggaca tgaccatgaa gtaccatatg gagggctcgg
     1081 tggacgggca caagttcgtg attacgggcc acgggaacgg caacccattc gaggggaaac
     1141 agacaatgaa cctgtgtgtg gtggaaggcg gacccctgcc cttctcggag gacatcctgt
     1201 ccgcaacctt tgactacggc aatcgcgtgt tcacagagta tccacagggt atggtggatt
     1261 tctttaagaa cagctgtccc gcaggataca cctggcaccg cagcctgctc ttcgaggacg
     1321 gggcggtgtg taccacctcc gctgatatca ccgtgtccgt cgaggagaac tgcttctacc
     1381 acaacagcaa attccacggc gtgaactttc ctgccgacgg gcctgtgatg aagaaaatga
     1441 ctacgaattg ggagcccagc tgcgagaaaa tcatcccggt ccctcgtcag ggaatcttga
     1501 agggcgacat cgcaatgtac ctcctgctca aggacggggg aagataccga tgccagttcg
     1561 ataccatcta caaggcaaag tccgatccca aggagatgcc agagtggcac ttcatccagc
     1621 acaaactcac ccgcgaggac cgctcagatg ccaagaacca gaagtggcaa ctggtcgagc
     1681 acgccgtggc atcccgtagc gccctgccag ggtaattcga atttaaatcg gatccctgtg
     1741 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1801 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1861 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1921 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg agatctgcgg ccgcgaagga
     1981 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2041 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaaaggccag gaaccgtaaa
     2101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3181 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3241 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3481 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3841 ccgcgcacat ttcccgccag atatacgcgt tgacattgat tattgactag ttattaatag
     3901 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3961 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     4021 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     4081 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     4141 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     4201 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     4261 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     4321 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     4381 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     4441 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     4501 aatacgactc actataggga gacccaagct tggtaccgag ctcggatcca ccggtcgcca
     4561 ccatgccggt ggacctcagc aagtggtccg ggcccttgag cctgcaagaa gtggacgagc
     4621 agccgcagca cccactgcat gtcacctacg ccggggcggc ggtggacgag ctgggcaaag
     4681 tgctgacgcc cacccaggtt aagaatagac ccaccagcat ttcgtgggat ggtcttgatt
     4741 cagggaagct ctacaccttg gtcctgacag acccggatgc tcccagcagg aaggatccca
     4801 aatacagaga atggcatcat ttcctggtgg tcaacatgaa gggcaatgac atcagcagtg
     4861 gcacagtcct ctccgattat gtgggctcgg ggcctcccaa gggcacaggc ctccaccgct
     4921 atgtctggct ggtttacgag caggacaggc cgctaaagtg tgacgagccc atcctcagca
     4981 accgatctgg agaccaccgt ggcaaattca aggtggcgtc cttccgtaaa aagtatgagc
     5041 tcagggcccc ggtggctggc acgtgttacc aggccgagtg ggatgactat gtgcccaaac
     5101 tgtacg
//
