LOCUS       sequence-322779-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for cloning user-defined spacers in the Tn6900 CRISPR 
            array..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Petassi MT, Hsieh SC, Peters JE
  TITLE     Guide RNA Categorization Enables Target Site Choice in 
            Tn7-CRISPR-Cas Transposons.
  JOURNAL   Cell. 2020 Nov 30. pii: S0092-8674(20)31464-1. doi: 
            10.1016/j.cell.2020.11.005.
  PUBMED    33271061
REFERENCE   2  (bases 1 to 5957)
  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: "Cell. 2020 
            Nov 30. pii: S0092-8674(20)31464-1. doi: 
            10.1016/j.cell.2020.11.005."
COMMENT     Sequence Label: pMTP170 (pBAD33_CRISPR(Tn6900)-entry)
FEATURES             Location/Qualifiers
     source          1..5957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      199..654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(286..305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     496..517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1210..1869)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1784..1803
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1870..1972)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2364..2381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2498..3043)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2578..2597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3119..3138
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3318..4196)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4223..4507
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4430..4449
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    4558..4572
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease 
                     (Haurwitz et al., 2010)"
     terminator      5383..5469
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5561..5588
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5607..5698
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggtgagcgt gggactcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
       61 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      121 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      181 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      241 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
      301 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
      361 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
      421 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
      481 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
      541 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
      601 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
      661 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      721 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      781 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      841 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      901 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      961 ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag aagcacacgg
     1021 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     1081 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     1141 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     1201 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     1261 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     1321 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     1381 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     1441 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     1501 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     1561 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     1621 caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc
     1681 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     1741 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     1801 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     1861 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     1921 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     1981 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     2041 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     2101 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     2161 ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg
     2221 ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     2281 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     2341 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     2401 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     2461 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     2521 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     2581 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     2641 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     2701 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     2761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     2821 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     2881 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     2941 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     3001 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     3061 tcagataaaa tatttctagg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     3121 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     3181 tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc
     3241 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt
     3301 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
     3361 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
     3421 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
     3481 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
     3541 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
     3601 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
     3661 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
     3721 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
     3781 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
     3841 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
     3901 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
     3961 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     4021 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     4081 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     4141 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     4201 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
     4261 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
     4321 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
     4381 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
     4441 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
     4501 tctccatacc cgtttttttg ggctagcgaa ttcgagctcg gtacccgggg atgtgaactg
     4561 ccgtataggc agccaagatt tgagacccac gatttgtaat aacagagtgt cttgtatttt
     4621 taaagaaagt ctatttaata caagtgatta tattaattaa cggtaagcat cagcgggtga
     4681 caaaacgagc atgcttacta ataaaatgtt aacctctgag gaagaattgt gaaactatca
     4741 ctaatggtag ctatatcgaa gaatggagtt atcgggaatg gccctgatat tccatggagt
     4801 gccaaaggtg aacagctcct gtttaaagct attacctata accaatggct gttggttgga
     4861 cgcaagactt ttgaatcaat gggagcatta cccaaccgaa agtatgcggt cgtaacacgt
     4921 tcaagtttta catctgacaa tgagaacgta ttgatctttc catcaattaa agatgcttta
     4981 accaacctaa agaaaataac ggatcatgtc attgtttcag gtggtgggga gatatacaaa
     5041 agcctgatcg atcaagtaga tacactacat atatctacaa tagacatcga gccggaaggt
     5101 gatgtttact ttcctgaaat ccccagcaat tttaggccag tttttaccca agacttcgcc
     5161 tctaacataa attatagtta ccaaatctgg caaaagggtt aacaagtggc agcaacggat
     5221 tcgcaaacct gtcacgcctt ttgtgccaaa agccgcgcca ggtttgcgat ccgctgtgcc
     5281 aggcgttagg cgtcatatga agatttcggt gatccctgag caggtctcat tttctgccgt
     5341 aaaggcagat attatttgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     5401 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5461 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     5521 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     5581 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta
     5641 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5701 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5761 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcagc
     5821 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5881 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5941 gctgataaat ctggagc
//
