LOCUS       sequence-412267-       10650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  R6K plasmid encoding a type V CAST but no CRISPR array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10650)
  TITLE     Distinct horizontal transfer mechanisms for type I and type V 
            CRISPR-associated transposons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10650)
  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            "Unpublished"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-CASTV
FEATURES             Location/Qualifiers
     source          1..10650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5507..5609
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5610..6269
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5676..5695)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    6286..6333
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(6411..6438)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6530..6616)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6917..6935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7003..7107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7108..7176,7177..7968)
                     /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(7326..7345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8377..8733
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            8784..8892
                     /note="incP origin of transfer"
     primer_bind     9133..9150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        9152..9186
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             9266..9277
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             10043..10054
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 cagtaaccat gtttggcaat gtgaccatac ccgcgtggat gtgttgctgg tagatcaaca
       61 tggtgaaatt ttaagtcgtc cctggctaac aacagtaatt gatacttact ctcgttgcat
      121 tatgggtatc aacttgggct ttgatgcacc cagttctggg gtagtagcat tagcgttacg
      181 ccatgcaatt ctaccaaagc gttacggttc cgagtacaaa ctgcattgtg agtggggaac
      241 ctatggaaaa ccagaacatt tttatactga tggcggtaaa gactttcgct ctaaccactt
      301 gagtcagatt ggggcgcaat tgggatttgt ctgtcattta cgcgatcgcc cttctgaagg
      361 tggagtagta gaacgtccct tcaaaacatt aaatgaccaa ctattttcaa cgcttcctgg
      421 gtacaccgga tctaatgtgc aggaacgccc agaagatgca gagaaggacg caagacttac
      481 tttgcgagaa ctagaacagt tacttgtgcg ttacatcgta gatcgttaca accaaagtat
      541 tgatgcgcgg atgggcgacc aaacgcgctt tgagcgttgg gaagcaggat tgcctacagt
      601 gccagtacca ataccagaac gagatttgga tatttgttta atgaagcagt cacggcgcac
      661 tgtgcaaaga ggtggttgtt tgcagtttca gaatttaatg tatcgggggg aatatttggc
      721 aggttatgcc ggagaaactg tcaacttaag gtttgacccc agagacatta caacaatttt
      781 ggtttatcgc caggaaaaca atcaggaagt atttctgact cgcgctcacg ctcaaggttt
      841 ggagacagag caactggcat tagatgaggc tgaggcagca agtcgcagac tccgtaccgc
      901 agggaaaact atcagtaacc aatcattatt gcaagaagtt gttgaccgcg atgctcttgt
      961 cgctaccaag aaaagccgta aggagcgtca aaaattggaa cagactgttt tgcgatctgc
     1021 tgctgttgat gaaagtaata gagaatcctt gccttctcaa atagttgaac cagatgaagt
     1081 ggaatctaca gaaacggttc actctcaata cgaagacatt gaggtgtggg actatgaaca
     1141 acttcgtgaa gaatatgggt tttaaacaat gacagaagct caggcgatcg ccaagcagtt
     1201 gggtggggta aaaccggatg atgagtggtt acaagctgaa attgctcgtc tcaagggtaa
     1261 gagcattgtg cctttacagc aggtaaaaac tctccatgat tggttagatg gcaagcgcaa
     1321 ggcaagaaaa tcttgccgag tagttgggga atcgagaact ggcaagacag ttgcttgtga
     1381 tgcctacaga tacaggcaca aacctcagca ggaagctgga cgacctccaa ctgtgcctgt
     1441 cgtttatatt cgacctcacc aaaaatgtgg ccccaaggat ttgtttaaaa agattactga
     1501 gtacctcaag tatcgggtaa caaaagggac tgtatctgat tttcgagata ggacgataga
     1561 agtactcaag ggttgtggcg tagagatgct aattattgat gaagctgacc gtctcaagcc
     1621 tgaaactttt gctgatgtgc gagatattgc cgaagattta ggaattgctg tggtactggt
     1681 aggaacagac cgtttggatg cggtaattaa gcgggatgag caggttctcg aacgctttcg
     1741 ggcgcatctt cgctttggta aattgtcggg agaggatttt aagaacaccg tagaaatgtg
     1801 ggaacaaatg gttttgaaac tgccagtatc ttctaatcta aagagcaagg agatgctacg
     1861 gattctcacg tcagcaactg aaggctacat tggtcgcctt gatgagattc ttagggaagc
     1921 tgcaattcgt tccttatcaa gaggattgaa gaagattgac aaggctgttt tacaggaagt
     1981 agctaaggag tacaaatgat agaagcacca gatgttaaac cttggctatt cttgattaaa
     2041 ccctatgaag gggaaagcct gagccacttt cttggcaggt tcagacgtgc caaccattta
     2101 tccgcaagtg gattgggtac tttggcagga attggtgcta tagtggcacg ttgggaaaga
     2161 tttcatttta atcctcgccc tagtcagcaa gaattggaag cgatcgcatc tgtagtagaa
     2221 gtggatgctc aaaggttagc ccagatgtta ccgcctgctg gagtgggaat gcagcatgag
     2281 ccaattcgct tgtgtggggc ttgttatgcc gagtcgcctt gtcaccgaat tgaatggcag
     2341 tacaagtcgg tgtggaagtg cgatcgccat caactcaaga ttttagcaaa gtgtccaaac
     2401 tgtcaagcac cttttaaaat gcctgcgctg tgggaggatg ggtgctgtca cagatgtagg
     2461 atgccgtttg cagaaatggc aaagctacag aaggtttgat gataaaacca gaaaaaggtg
     2521 tgaaattaac taagtccctg aattgatctg gttgtccaaa aaatttgtgc gatcgcatgg
     2581 caagattatt cctactattg atgtggcgtg agtcggataa cttgctctaa tgggatcccg
     2641 gttaatacca atagatcaga tatgagtaat aaacattacg acctttacct gtggtattca
     2701 ctgttggcac gacaacacct ttatcgcgcc aatactgtag ctgacgacgc gaacaattag
     2761 tgatttgagc cgcttctgtg cttgtgtaga attttccttc catagaaatc atcctacaga
     2821 agtagttttt taaatttaca ttttttaaaa gcacattatg tattcatgtg atattatttt
     2881 tttatgagtc aaataactat tcaagctcga cttatttcct ttgaatcaaa ccgccaacaa
     2941 ctctggaagt tgatggcaga tttaaacacg ccgttaatta acgaactgct ttgccagtta
     3001 ggtcaacacc ccgacttcga gaagtggcaa caaaagggta aactcccgtc taccgttgtg
     3061 agccagttat gtcaacctct caaaactgac cctcgctttg caggtcagcc cagccgttta
     3121 tatatgtcgg caattcatat tgtggactac atctacaagt cctggctggc tatacagaaa
     3181 cggcttcaac agcagctaga tggaaagacg cgctggctag aaatgctcaa tagcgatgct
     3241 gaattagtag aacttagtgg tgacacttta gaggctattc gtgtcaaagc tgctgaaatt
     3301 ttggcaatag ctatgccagc atctgagtca gatagcgctt cacctaaagg gaaaaaaggt
     3361 aaaaaggaga aaaaaccctc atcttctagc cctaagcgta gtttatccaa gacattattt
     3421 gacgcttacc aagaaacgga agatatcaag agccgtagcg ccatcagcta cctgttaaaa
     3481 aatggctgca aacttactga caaagaagaa gattcagaaa aatttgctaa acgtcgtcgt
     3541 caagttgaaa tccaaattca aaggcttacc gaaaagttaa taagtcggat gcctaaaggt
     3601 cgagatttga ccaatgctaa atggttggag acactcttga ctgctacaac cactgttgct
     3661 gaagacaacg cccaagccaa acgctggcag gatattctgt taactcgatc aagttctctc
     3721 ccattccccc ttgtttttga aaccaacgag gatatggttt ggtcaaagaa tcaaaagggt
     3781 aggctgtgtg ttcacttcaa tggcttaagc gatttaattt ttgaggtgta ctgcggcaat
     3841 cgtcaacttc actggtttca acgcttccta gaagaccaac agactaaacg caaaagcaaa
     3901 aatcagcatt ctagcggctt gttcacactc agaaatggtc atctagtttg gcttgaaggt
     3961 gagggtaaag gggaaccttg gaatcttcac cacttgaccc tttactgctg tgttgacaat
     4021 cgcttgtgga cagaggaggg aacagaaatc gttcgccaag agaaagcaga tgaaattact
     4081 aaattcatca caaacatgaa gaagaaaagc gatctaagcg atacacagca agctttgatt
     4141 caacgtaaac aatcaacact tactcgaata aacaattcct ttgagcgtcc tagccaaccc
     4201 ctttatcaag gtcaatcaca cattttggtt ggagtaagcc tgggactaga aaaacctgcc
     4261 acagtagcag tagtagatgc gatcgccaac aaagtcttgg cttaccggag tattaaacaa
     4321 ttacttggcg acaattacga actgctaaat cgccagagac gacaacagca gtacctatct
     4381 cacgaacgcc acaaagcaca aaaaaacttc tctcccaatc aatttggagc atctgagtta
     4441 gggcaacata tagacagatt attagctaaa gcaattgtag cgttagcgag aacctacaaa
     4501 gctggcagta ttgtcttgcc caagttaggg gatatgcggg aggttgtcca aagtgaaatt
     4561 caagctatag cagaacaaaa atttcccggt tatattgaag gtcagcaaaa atatgccaaa
     4621 cagtaccggg ttaatgttca tcggtggagc tacggcagat taattcaaag cattcaaagt
     4681 aaagcagctc aaacaggaat tgtgattgag gagggaaaac aacctattcg aggtagtccc
     4741 cacgacaaag caaaggaatt agcactttct gcttacaatc tccgcctaac taggcgaagt
     4801 taacaaatat ctgaaccttg ataatagaat attaatagcg ccgcaattca tgctgcttgc
     4861 agcctctgaa ttttgttaaa tgagggttag tttgactgta taaatacagt cttgctttct
     4921 gaccctggta gctgctcacc ctgatgctgc tgtcaataga caggataggt gcgctcccag
     4981 caataagggc gcggatgtac tgctgtagtg gctactgaat cacccccgat caagggggaa
     5041 ccctccccaa ttcttcattt gaaggactaa aatcaaggca aaatttctaa gagatccgcg
     5101 caagttccaa ataccttatc tcgtcttgat ttcatctttt tttaaccaag gcatgattct
     5161 tgagactgag gctcaaatga gaaattggga aacatccgcg ctgaagatat ctggaaaggt
     5221 tggctcacaa cattttgtaa ctgggtagaa cgcaaaacca ttgtagagag gtagcattgc
     5281 tacgtctcta cgtgtacagt gactaattat atgtcgttgt gacaaattat tgtcatcagt
     5341 aaaatcctta tacagtatag attatagcgc tttggcagtt ttagcataac ctctttgcag
     5401 tgacaaaata gatgtcgttg tccgtgattg tgacaaatta gctgtcgctt tgcaagatag
     5461 gaaaaagctt ttgtgtattt tcataatgac aaattgactg tcgccttgat cggcacgtaa
     5521 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttgt
     5581 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     5641 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     5701 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     5761 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     5821 cggaattacg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     5881 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     5941 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     6001 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
     6061 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     6121 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     6181 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     6241 actgcgatga gtggcagggc ggggcgtaag gcgcgccatt taaatgaagt tcctattccg
     6301 aagttcctat tctctagaaa gtataggaac ttcgaagcag ctccagccta cacaatcgct
     6361 caagacgtgt aatgctgcaa tctgcatgca agcttggcac tggccacgca aaaaggccat
     6421 ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg cgtcctgccc
     6481 gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga tttgtcctac
     6541 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     6601 cctttcattt tatttgtcag taactattaa acttaggggt gggttgaaag caagtccttt
     6661 tatccgcttg ttttaattgc tttgtataat aattgcagag catattatat tgatgacatt
     6721 taatttgtca tcaattaatt aagcaacgct gatgggtcac gacgacaatt aaatagtcac
     6781 aatgacatta atctgtcacc gacgacagat aatttgtcac tgtacactac gccttttgtg
     6841 gagatgtcta ataaactaaa aattttttat gatcagaggt tttcaccgtc atcaccgaaa
     6901 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6961 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     7021 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     7081 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     7141 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     7201 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     7261 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     7321 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     7381 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     7441 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7501 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     7561 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7621 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7681 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7741 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7801 aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt
     7861 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7921 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7981 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     8041 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     8101 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     8161 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8221 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8281 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8341 acatacctcg ctctgctaat cctgttacca gccggttgtc agccgttaag tgttcctgtg
     8401 tcactcaaaa ttgctttgag aggctctaag ggcttctcag tgcgttacat ccctggcttg
     8461 ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt atatattctt ttttttctta
     8521 taaaacttaa aaccttagag gctatttaag ttgctgattt atattaattt tattgttcaa
     8581 acatgagagc ttagtacgtg aaacatgaga gcttagtacg ttagccatga gagcttagta
     8641 cgttagccat gagggtttag ttcgttaaac atgagagctt agtacgttaa acttgagagc
     8701 ttagtacgtg aaacatgaga gcttagtacg tactatcaac aggttgaact gcccatgttc
     8761 tttcctgcgt tatcagagct tatcggccag cctcgcagag caggattccc gttgagcacc
     8821 gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca
     8881 cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg
     8941 caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc
     9001 ccgaagcagg gttatgcagc ggaaagtata ccttaaatgt tctttcctgc gttatcccct
     9061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     9121 acgaccgagc gcagcgagtc agtgagcgag gttgacagct agctcagtcc taggtataat
     9181 actagcgtcg acgtggagat ataccaaacg agagacccat cgcggcaact caagaaacgg
     9241 caggttcgga tcttagctac tagagaaaga ggagaaatac tagatgcgta aaggcgaaga
     9301 gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca acggtcataa
     9361 gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt
     9421 catctgtact actggtaaac tgccggttcc ttggccgact ctggtaacga cgctgactta
     9481 tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact tcttcaagtc
     9541 cgccatgccg gaaggctatg tgcaggaacg cacgatttga cggcacgtac aaaacgcgtg
     9601 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     9661 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     9721 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     9781 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     9841 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     9901 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     9961 cgcatggtat ggatgaactg tacaaatgac caggcatggt ctcctatgga tccgaattcg
    10021 agcgaaggag atatacatcc ctaaagagga gaaaggatct atgaacagtc agcaaaatcc
    10081 tgatttagct gttcatccct tggcaattcc tatggaaggc ttactaggag aaagtgctac
    10141 aactcttgag aagaatgtaa ttgccacaca actctcagag gaagcccaag taaagctaga
    10201 ggtaatccaa agtttactgg aaccctgcga tcgcacaact tatgggcaaa agttgcggga
    10261 agcagcagag aaactaaatg tatcgttgcg aacggtacaa aggttggtga aaaactggga
    10321 acaagatggc ttagtcggac tcactcaaac aagtagggct gataaaggaa aacaccgcat
    10381 tggtgagttt tgggaaaact tcattaccaa aacctacaag gagggtaaca agggaagtaa
    10441 acgtatgacc cctaaacaag ttgctctcag agtcgaggct aaagcccgtg aattaaaaga
    10501 ctctaagccg cccaattaca aaaccgtgtt acgggtatta gcacccattt tggaaaagca
    10561 acaaaaagcc aagagtatcc gcagtcctgg ttggagagga actacgcttt cggttaaaac
    10621 ccgtgaagga aaagatttat cggttgatta
//
