LOCUS       sequence-412250-        4394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  R6K backbone with Golden Gate (GG) cloning sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4394)
  TITLE     Distinct horizontal transfer mechanisms for type I and type V 
            CRISPR-associated transposons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4394)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pR6K-GFP-RFP
FEATURES             Location/Qualifiers
     source          1..4394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1171..1527
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            1578..1686
                     /note="incP origin of transfer"
     primer_bind     1927..1944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1946..1980
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             2060..2071
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2855..2863
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2869..3564
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3002..3020)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3270..3289
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3568..3639
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3655..3682
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3701..3718)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3701..3717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3710..3732)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3926..3945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4045..4067
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4105..4123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4191..4295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4296..4364,4365..4394,1..762)
                     /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"
ORIGIN
        1 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
       61 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
      181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
      241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
      301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
      361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
      421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
      481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
      541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
      601 ggagccggtg agcgtggctc tcgcggtatc attgcagcac tggggccaga tggtaagccc
      661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
      721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
      781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
      841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
      901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1141 ctcgctctgc taatcctgtt accagccggt tgtcagccgt taagtgttcc tgtgtcactc
     1201 aaaattgctt tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc
     1261 acaaccgtta aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac
     1321 ttaaaacctt agaggctatt taagttgctg atttatatta attttattgt tcaaacatga
     1381 gagcttagta cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag
     1441 ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacttga gagcttagta
     1501 cgtgaaacat gagagcttag tacgtactat caacaggttg aactgcccat gttctttcct
     1561 gcgttatcag agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     1621 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     1681 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     1741 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     1801 cagggttatg cagcggaaag tataccttaa atgttctttc ctgcgttatc ccctgattct
     1861 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1921 gagcgcagcg agtcagtgag cgaggttgac agctagctca gtcctaggta taatactagc
     1981 gtcgacgtgg agatatacca aacgagagac ccatcgcggc aactcaagaa acggcaggtt
     2041 cggatcttag ctactagaga aagaggagaa atactagatg cgtaaaggcg aagagctgtt
     2101 cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc
     2161 cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg
     2221 tactactggt aaactgccgg ttccttggcc gactctggta acgacgctga cttatggtgt
     2281 tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat
     2341 gccggaaggc tatgtgcagg aacgcacgat ttgacggcac gtacaaaacg cgtgcggaag
     2401 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     2461 aggacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     2521 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     2581 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     2641 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     2701 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     2761 gtatggatga actgtacaaa tgaccaggca tggtctccta tggatccgaa ttcgagcgaa
     2821 ggagatatac atccctttaa gcaggtgttt caattaagga ggttttttat ggtgagcaag
     2881 ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac
     2941 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3001 gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct
     3061 cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc cgactactat
     3121 aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3181 gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag
     3241 ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac aatgggctgg
     3301 gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg
     3361 gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac ctacaaggcc
     3421 aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc
     3481 cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc
     3541 ggcggcatgg acgagctgta caagtgacaa ataaaacgaa aggctcagtc gaaagactgg
     3601 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     3661 ccttcgggtg ggcctttctg cgcacctgca tgctatgatc actggccgtc gttttacaac
     3721 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3781 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3841 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3901 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3961 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4021 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4081 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4141 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4201 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4261 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4321 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4381 tgaaagtaaa agat
//
